DOI: 10.1126/science.279.5351.703. Transcription Factor-Specific Requirements for Coactivators and Their Acetyltransferase Functions

Transcription Factor-Specific Requirements for Coactivators and Their Acetyltransferase Functions

10.1126/science.279.5351.703

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

Different classes of mammalian transcription factors—nuclear receptors, cyclic adenosine 3′,5′-monophosphate–regulated enhancer binding protein (CREB), and signal transducer and activator of transcription-1 (STAT-1)—functionally require distinct components of the coactivator complex, including CREB-binding protein (CBP/p300), nuclear receptor coactivators (NCoAs), and p300/CBP-associated factor (p/CAF), based on their platform or assembly properties. Retinoic acid receptor, CREB, and STAT-1 also require different histone acetyltransferase (HAT) activities to activate transcription. Thus, transcription factor–specific differences in configuration and content of the coactivator complex dictate requirements for specific acetyltransferase activities, providing an explanation, at least in part, for the presence of multiple HAT components of the complex.

Bibliography

Korzus, E., Torchia, J., Rose, D. W., Xu, L., Kurokawa, R., McInerney, E. M., Mullen, T.-M., Glass, C. K., & Rosenfeld, M. G. (1998). Transcription Factor-Specific Requirements for Coactivators and Their Acetyltransferase Functions. Science, 279(5351), 703â707.

Authors 9

Edward Korzus (first)
Joseph Torchia (additional)
David W. Rose (additional)
Lan Xu (additional)
Riki Kurokawa (additional)
Eileen M. McInerney (additional)
Tina-Marie Mullen (additional)
Christopher K. Glass (additional)
Michael G. Rosenfeld (additional)

References 61 Referenced 493

10.1038/365855a0
Eckner R., et al., Genes Dev. 8, 869 (1994); (10.1101/gad.8.8.869) / Genes Dev. by Eckner R. (1994)
Kwok R. P., et al., Nature 370, 223 (1994). (10.1038/370223a0) / Nature by Kwok R. P. (1994)
Kamei Y., et al., Cell 85, 403 (1996). (10.1016/S0092-8674(00)81118-6) / Cell by Kamei Y. (1996)
Chakravarti D., et al., Nature 383, 99 (1996); (10.1038/383099a0) / Nature by Chakravarti D. (1996)
Hanstein B., et al., Proc. Natl. Acad. Sci. U.S.A. 93, 11540 (1996); (10.1073/pnas.93.21.11540) / Proc. Natl. Acad. Sci. U.S.A. by Hanstein B. (1996)
; T. P. Yao G. Ku N. Zhou R. Scully D. M. Livingston ibid. p. 10626.
10.1126/science.8197458
Cavailles V., Dauvois S., Danielian P. S., Parker M. G., Proc. Natl. Acad. Sci. U.S.A. 91, 10009 (1994). (10.1073/pnas.91.21.10009) / Proc. Natl. Acad. Sci. U.S.A. by Cavailles V. (1994)
S. A. Onate S. Y. Tsai M.-J. Tsai B. W. O'Malley Science 270 1354 (1995). (10.1126/science.270.5240.1354)
Voegel J. J., Heine M. J., Zechel C., Chambon P., Gronemeyer H., EMBO J. 15, 3667 (1996); (10.1002/j.1460-2075.1996.tb00736.x) / EMBO J. by Voegel J. J. (1996)
Hong H., Kohli K., Garabedian M. J., Stallcup M. R., Mol. Cell. Biol. 17, 2735 (1997). (10.1128/MCB.17.5.2735) / Mol. Cell. Biol. by Hong H. (1997)
10.1038/42652
A human homolog of mouse p/CIP gene has been also described [
10.1016/S0092-8674(00)80516-4
Li H., Gomes P. J., Chen J. D., Proc. Natl. Acad. Sci. U.S.A. 94, 8479 (1997); (10.1073/pnas.94.16.8479) / Proc. Natl. Acad. Sci. U.S.A. by Li H. (1997)
; O. M. Sobulo et al. ibid. p. 8732].
Yang X. J., Ogryzko V. V., Nishikawa J., Howard B. H., Nakatani Y., Nature 382, 319 (1996). (10.1038/382319a0) / Nature by Yang X. J. (1996)
Marcus G. A., Silverman N., Berger S. L., Horiuchi J., Guarente L., EMBO J. 13, 4807 (1994); (10.1002/j.1460-2075.1994.tb06806.x) / EMBO J. by Marcus G. A. (1994)
Candau R., et al., Mol. Cell. Biol. 16, 593 (1996); (10.1128/MCB.16.2.593) / Mol. Cell. Biol. by Candau R. (1996)
Candau R., Berger S. L., J. Biol. Chem. 271, 5237 (1996); (10.1074/jbc.271.9.5237) / J. Biol. Chem. by Candau R. (1996)
10.1016/S0092-8674(00)81063-6
Candau R., Zhou J. X., Allis C. D., Berger S. L., EMBO J. 16, 555 (1997). (10.1093/emboj/16.3.555) / EMBO J. by Candau R. (1997)
Ogryzko V. V., Schiltz R. L., Russanova V., Howard B. H., Nakatani Y., Cell 87, 953 (1996); (10.1016/S0092-8674(00)82001-2) / Cell by Ogryzko V. V. (1996)
Bannister A. J., Kouzarides T., Nature 384, 641 (1996). (10.1038/384641a0) / Nature by Bannister A. J. (1996)
Spencer T. E., et al., Nature 389, 194 (1997). (10.1038/38304) / Nature by Spencer T. E. (1997)
Heery D. M., Kalkhoven E., Hoare S., Parker M., ibid. 387, 733 (1997). / ibid. by Heery D. M. (1997)
Glass C. K., Lipkin S. M., Devary O. V., Rosenfeld M. G., Cell 59, 697 (1989); (10.1016/0092-8674(89)90016-0) / Cell by Glass C. K. (1989)
Kurokawa R., et al., Nature 377, 451 (1995). (10.1038/377451a0) / Nature by Kurokawa R. (1995)
Horlein A. J., et al., Nature 377, 397 (1995). (10.1038/377397a0) / Nature by Horlein A. J. (1995)
Alland L., et al., ibid. 387, 49 (1997); / ibid. by Alland L. (1997)
Hassig C. A., Fleischer T. C., Billin A. N., Schreiber S. L., Ayer D. E., Cell 89, 341 (1997); (10.1016/S0092-8674(00)80214-7) / Cell by Hassig C. A. (1997)
Heinzel T., et al., Nature 387, 43 (1997); (10.1038/387043a0) / Nature by Heinzel T. (1997)
10.1016/S0092-8674(00)80217-2
; C. D. Laherty et al. ibid. p. 349; L. Nagy et al. ibid. p. 373; Y. Zhang R. Iratni H. Erdjument-Bromage P. Tempst D. Reinberg ibid. p. 357.
Danielian P. S., White R., Lees J. A., Parker M. G., EMBO J. 11, 1025 (1992); (10.1002/j.1460-2075.1992.tb05141.x) / EMBO J. by Danielian P. S. (1992)
Durand B., et al., ibid. 13, 5370 (1994). / ibid. by Durand B. (1994)
Berger S. L., et al., Cell 70, 251 (1992); (10.1016/0092-8674(92)90100-Q) / Cell by Berger S. L. (1992)
Georgakopoulos T., Thireos G., EMBO J. 11, 4145 (1992). (10.1002/j.1460-2075.1992.tb05507.x) / EMBO J. by Georgakopoulos T. (1992)
Swope D. L., Mueller C. L., Chrivia J. C., J. Biol. Chem. 271, 28138 (1996); (10.1074/jbc.271.45.28138) / J. Biol. Chem. by Swope D. L. (1996)
; S. Bisotto S. Minorgan R. P. Rehfuss ibid. p. 17746.
E. Korzus D. W. Rose M. G. Rosenfeld data not shown.
Neuwald A. F., Landsman D., Trends Biochem. Sci. 22, 154 (1997). (10.1016/S0968-0004(97)01034-7) / Trends Biochem. Sci. by Neuwald A. F. (1997)
10.1126/science.279.5351.700
Zhang J. J., et al., Proc. Natl. Acad. Sci. U.S.A. 93, 15092 (1996); (10.1073/pnas.93.26.15092) / Proc. Natl. Acad. Sci. U.S.A. by Zhang J. J. (1996)
Bhattacharya S., et al., Nature 383, 344 (1996); (10.1038/383344a0) / Nature by Bhattacharya S. (1996)
Horvai A. E., et al., Proc. Natl. Acad. Sci. U.S.A. 94, 1074 (1997). (10.1073/pnas.94.4.1074) / Proc. Natl. Acad. Sci. U.S.A. by Horvai A. E. (1997)
10.1126/science.277.5332.1630
Whole-cell extracts were prepared by lysing the cells in NET-N buffer containing 50 mM tris (pH 7.6) 5 mM EDTA 0.3 M NaCl 1 mM dithiothreitol 0.1% NP-40 and protease inhibitors (2 mM phenylmethylsulfonyl fluoride and 10 μg/ml each of leupeptin pepstatin and aprotinin). The mixture was then centrifuged at 30 000 g for 1 hour at 4°C and the supernatant was stored at –80°C until use. For coimmunoprecipitation assays 1 mg of cell extract was incubated in the presence of 2 μg of anti-p/CIP anti-Flag or anti-RAR IgG for 2 hours at 4°C. The immune complexes were then precipitated with protein A–Sepharose. Protein complexes were resolved by SDS–polyacrylamide gel electrophoresis (PAGE) [
10.1038/227680a0
] protein immunoblotted and probed using 1 μg/ml of anti-Flag anti-p/CIP or an anti-hemagglutinin (anti-HA).
The yeast strain EGY 48 the LexA-b galactosidase reporter construct (PSH 18-34) and the B42 parental vectors (PEG 202 and PJG 4-5) were all previously described [
10.1016/0092-8674(93)90498-F
]. Various p/CAF fragments or fragments of other coactivators were obtained by polymerase chain reaction or restriction digestion and subcloned into PEG 202 bait vector or PJG 4-5 prey vectors respectively. EGY 48 cells were transformed with the lac Z reporter plasmid pSH 18-34 with the appropriate bait and prey vectors and were plated out on Ura – His – Trp – medium containing 2% galactose. Isolated yeast colonies were then allowed to grow in the same liquid medium and were subsequently assayed for β-galactosidase as previously described [F. M. Ausubel et al. Current Protocols in Molecular Biology (Wiley New York 1995)]. All constructs were sequenced and functionally tested in yeast two-hybrid assays showing interaction with other proteins as previously described (2 7). In the case of p/CAF constructs each fragment interacted positively with at least one other coactivator except the fragment with amino acids 86 through 518.
Insulin-responsive Rat-1 fibroblasts were seeded on acid-washed glass cover slips at subconfluent density and grown in D-MEM/F-12 (Life Technologies) medium supplemented with 10% fetal bovine serum gentamicin and methotrexate. Before the injection the cells were rendered quiescent by incubation in serum-free medium for 24 to 36 hours. Plasmids were injected into the nuclei of cells at a final concentration of 100 μg/ml. Immunoglobulin G specific for p/CAF was prepared from guinea pig serum raised against a bacterially expressed fragment (amino acids 466 to 832) of p/CAF; this IgG recognized the single band of p/CAF in protein immunoblot analysis. Either preimmune IgG or the appropriate specific antibodies directed against p/CAF p/CIP SRC-1 or CBP were coinjected and allowed the unambiguous identification of the injected cells (7). Preimmune controls were included in all experiments. Microinjections were carried out using an Eppendorf semiautomated microinjection system mounted on an inverted Zeiss microscope. Approximately 1 hour after injection the cells were stimulated where indicated with the appropriate ligand. In the case of rescue experiments the cells were stimulated with ligand 6 hours after injection to allow protein expression. After overnight incubation the cells were fixed and then stained to detect injected IgG and β-galactosidase expression [
Rose D. W., et al., J. Cell Biol. 119, 1405 (1992); (10.1083/jcb.119.6.1405) / J. Cell Biol. by Rose D. W. (1992)
; (2)]. Injected cells were identified by staining with tetramethylrhodamine-conjugated donkey anti-rabbit IgG. All experimental results are expressed as the mean ± SEM of at least three experiments in which at least 1000 cells were injected.
GST-RAR and GST-CBP fragments were generated as described (2). We incubated 25 μl of GST-Sepharose beads containing 3 to 6 μg of the GST recombinant proteins with 5 × 10 5 cpm of 35 S-labeled p/CAF proteins generated by in vitro transcription and translation for 2 hours at 4°C. The complexes were washed five times with NET-N buffer resolved by SDS-PAGE and fluorographed.
Mutations in p/CAF and CBP were introduced by site-directed mutagenesis using the Quick-Change mutagenesis system (Stratagene) according to the manufacturer's instructions. Double-stranded oligonucleotides were designed such that the wild-type sequence corresponding to amino acids Tyr 616 /Phe 617 (acetyl-CoA–binding site) in p/CAF cDNA were substituted with alanines in order to generate a mutant of p/CAF lacking HAT activity (pCMV– p/CAF HAT– ). A similar strategy was used to obtain mutants of CBP. Mutants of p/CAF and CBP were expressed in bacteria and baculovirus respectively and tested for HAT activity in solution using histones as substrates [
10.1073/pnas.92.14.6364
We thank R. Heyman for use of TTNPB and LG629 S. L. Berger for discussion and providing hGCN5 expression vector and Y. Nakatani and X. J. Yang for providing a tagged p/CAF expression vector. We also thank C. Nelson for experimental assistance L.-M. Phillips for excellent technical assistance and P. Myer for expertise in figure preparation. E.K. is supported by a U.S. Army Medical Research Program Award J.T. by the National Cancer Institute of Canada E.M.M. by an NIH Postdoctoral Fellowship D.W.R. by an American Diabetes Association Career Development Award and L.X. by an American Heart Association Predoctoral Fellowship. Supported by grants from NIH to C.K.G. and M.G.R.

Dates

Type	When
Created	23 years ago (July 27, 2002, 5:42 a.m.)
Deposited	1 year, 7 months ago (Jan. 13, 2024, 12:58 a.m.)
Indexed	1 month, 3 weeks ago (July 2, 2025, 3:07 p.m.)
Issued	27 years, 6 months ago (Jan. 30, 1998)
Published	27 years, 6 months ago (Jan. 30, 1998)
Published Print	27 years, 6 months ago (Jan. 30, 1998)

Funders 0

None

BibTeX

@article{Korzus_1998, title={Transcription Factor-Specific Requirements for Coactivators and Their Acetyltransferase Functions}, volume={279}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.279.5351.703}, DOI={10.1126/science.279.5351.703}, number={5351}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Korzus, Edward and Torchia, Joseph and Rose, David W. and Xu, Lan and Kurokawa, Riki and McInerney, Eileen M. and Mullen, Tina-Marie and Glass, Christopher K. and Rosenfeld, Michael G.}, year={1998}, month=jan, pages={703–707} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        7,
        2
      ]
    ],
    "date-time": "2025-07-02T19:07:47Z",
    "timestamp": 1751483267035
  },
  "reference-count": 61,
  "publisher": "American Association for the Advancement of Science (AAAS)",
  "issue": "5351",
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        1998,
        1,
        30
      ]
    ]
  },
  "abstract": "<jats:p>Different classes of mammalian transcription factors\u2014nuclear receptors, cyclic adenosine 3\u2032,5\u2032-monophosphate\u2013regulated enhancer binding protein (CREB), and signal transducer and activator of transcription-1 (STAT-1)\u2014functionally require distinct components of the coactivator complex, including CREB-binding protein (CBP/p300), nuclear receptor coactivators (NCoAs), and p300/CBP-associated factor (p/CAF), based on their platform or assembly properties. Retinoic acid receptor, CREB, and STAT-1 also require different histone acetyltransferase (HAT) activities to activate transcription. Thus, transcription factor\u2013specific differences in configuration and content of the coactivator complex dictate requirements for specific acetyltransferase activities, providing an explanation, at least in part, for the presence of multiple HAT components of the complex.</jats:p>",
  "DOI": "10.1126/science.279.5351.703",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2002,
        7,
        27
      ]
    ],
    "date-time": "2002-07-27T09:42:47Z",
    "timestamp": 1027762967000
  },
  "page": "703-707",
  "source": "Crossref",
  "is-referenced-by-count": 493,
  "title": "Transcription Factor-Specific Requirements for Coactivators and Their Acetyltransferase Functions",
  "prefix": "10.1126",
  "volume": "279",
  "author": [
    {
      "given": "Edward",
      "family": "Korzus",
      "sequence": "first",
      "affiliation": [
        {
          "name": "E. Korzus, J. Torchia, E. M. McInerney, M. G. Rosenfeld, Howard Hughes Medical Institute, University of California San Diego (UCSD), 9500 Gilman Drive, La Jolla, CA 92093\u20130648, USA."
        },
        {
          "name": "D. W. Rose and T.-M. Mullen, UCSD Department of Medicine and Whittier Diabetes Program, UCSD."
        },
        {
          "name": "L. Xu, Howard Hughes Medical Institute and Biomedical Sciences Graduate Program, UCSD."
        },
        {
          "name": "R. Kurokawa and C. K. Glass, Cellular and Molecular Medicine, School and Department of Medicine, UCSD."
        }
      ]
    },
    {
      "given": "Joseph",
      "family": "Torchia",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "E. Korzus, J. Torchia, E. M. McInerney, M. G. Rosenfeld, Howard Hughes Medical Institute, University of California San Diego (UCSD), 9500 Gilman Drive, La Jolla, CA 92093\u20130648, USA."
        },
        {
          "name": "D. W. Rose and T.-M. Mullen, UCSD Department of Medicine and Whittier Diabetes Program, UCSD."
        },
        {
          "name": "L. Xu, Howard Hughes Medical Institute and Biomedical Sciences Graduate Program, UCSD."
        },
        {
          "name": "R. Kurokawa and C. K. Glass, Cellular and Molecular Medicine, School and Department of Medicine, UCSD."
        }
      ]
    },
    {
      "given": "David W.",
      "family": "Rose",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "E. Korzus, J. Torchia, E. M. McInerney, M. G. Rosenfeld, Howard Hughes Medical Institute, University of California San Diego (UCSD), 9500 Gilman Drive, La Jolla, CA 92093\u20130648, USA."
        },
        {
          "name": "D. W. Rose and T.-M. Mullen, UCSD Department of Medicine and Whittier Diabetes Program, UCSD."
        },
        {
          "name": "L. Xu, Howard Hughes Medical Institute and Biomedical Sciences Graduate Program, UCSD."
        },
        {
          "name": "R. Kurokawa and C. K. Glass, Cellular and Molecular Medicine, School and Department of Medicine, UCSD."
        }
      ]
    },
    {
      "given": "Lan",
      "family": "Xu",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "E. Korzus, J. Torchia, E. M. McInerney, M. G. Rosenfeld, Howard Hughes Medical Institute, University of California San Diego (UCSD), 9500 Gilman Drive, La Jolla, CA 92093\u20130648, USA."
        },
        {
          "name": "D. W. Rose and T.-M. Mullen, UCSD Department of Medicine and Whittier Diabetes Program, UCSD."
        },
        {
          "name": "L. Xu, Howard Hughes Medical Institute and Biomedical Sciences Graduate Program, UCSD."
        },
        {
          "name": "R. Kurokawa and C. K. Glass, Cellular and Molecular Medicine, School and Department of Medicine, UCSD."
        }
      ]
    },
    {
      "given": "Riki",
      "family": "Kurokawa",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "E. Korzus, J. Torchia, E. M. McInerney, M. G. Rosenfeld, Howard Hughes Medical Institute, University of California San Diego (UCSD), 9500 Gilman Drive, La Jolla, CA 92093\u20130648, USA."
        },
        {
          "name": "D. W. Rose and T.-M. Mullen, UCSD Department of Medicine and Whittier Diabetes Program, UCSD."
        },
        {
          "name": "L. Xu, Howard Hughes Medical Institute and Biomedical Sciences Graduate Program, UCSD."
        },
        {
          "name": "R. Kurokawa and C. K. Glass, Cellular and Molecular Medicine, School and Department of Medicine, UCSD."
        }
      ]
    },
    {
      "given": "Eileen M.",
      "family": "McInerney",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "E. Korzus, J. Torchia, E. M. McInerney, M. G. Rosenfeld, Howard Hughes Medical Institute, University of California San Diego (UCSD), 9500 Gilman Drive, La Jolla, CA 92093\u20130648, USA."
        },
        {
          "name": "D. W. Rose and T.-M. Mullen, UCSD Department of Medicine and Whittier Diabetes Program, UCSD."
        },
        {
          "name": "L. Xu, Howard Hughes Medical Institute and Biomedical Sciences Graduate Program, UCSD."
        },
        {
          "name": "R. Kurokawa and C. K. Glass, Cellular and Molecular Medicine, School and Department of Medicine, UCSD."
        }
      ]
    },
    {
      "given": "Tina-Marie",
      "family": "Mullen",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "E. Korzus, J. Torchia, E. M. McInerney, M. G. Rosenfeld, Howard Hughes Medical Institute, University of California San Diego (UCSD), 9500 Gilman Drive, La Jolla, CA 92093\u20130648, USA."
        },
        {
          "name": "D. W. Rose and T.-M. Mullen, UCSD Department of Medicine and Whittier Diabetes Program, UCSD."
        },
        {
          "name": "L. Xu, Howard Hughes Medical Institute and Biomedical Sciences Graduate Program, UCSD."
        },
        {
          "name": "R. Kurokawa and C. K. Glass, Cellular and Molecular Medicine, School and Department of Medicine, UCSD."
        }
      ]
    },
    {
      "given": "Christopher K.",
      "family": "Glass",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "E. Korzus, J. Torchia, E. M. McInerney, M. G. Rosenfeld, Howard Hughes Medical Institute, University of California San Diego (UCSD), 9500 Gilman Drive, La Jolla, CA 92093\u20130648, USA."
        },
        {
          "name": "D. W. Rose and T.-M. Mullen, UCSD Department of Medicine and Whittier Diabetes Program, UCSD."
        },
        {
          "name": "L. Xu, Howard Hughes Medical Institute and Biomedical Sciences Graduate Program, UCSD."
        },
        {
          "name": "R. Kurokawa and C. K. Glass, Cellular and Molecular Medicine, School and Department of Medicine, UCSD."
        }
      ]
    },
    {
      "given": "Michael G.",
      "family": "Rosenfeld",
      "sequence": "additional",
      "affiliation": [
        {
          "name": "E. Korzus, J. Torchia, E. M. McInerney, M. G. Rosenfeld, Howard Hughes Medical Institute, University of California San Diego (UCSD), 9500 Gilman Drive, La Jolla, CA 92093\u20130648, USA."
        },
        {
          "name": "D. W. Rose and T.-M. Mullen, UCSD Department of Medicine and Whittier Diabetes Program, UCSD."
        },
        {
          "name": "L. Xu, Howard Hughes Medical Institute and Biomedical Sciences Graduate Program, UCSD."
        },
        {
          "name": "R. Kurokawa and C. K. Glass, Cellular and Molecular Medicine, School and Department of Medicine, UCSD."
        }
      ]
    }
  ],
  "member": "221",
  "reference": [
    {
      "key": "e_1_3_1_2_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/365855a0"
    },
    {
      "key": "e_1_3_1_2_3",
      "doi-asserted-by": "crossref",
      "first-page": "869",
      "DOI": "10.1101/gad.8.8.869",
      "volume": "8",
      "author": "Eckner R.",
      "year": "1994",
      "unstructured": "Eckner R., et al., Genes Dev. 8, 869 (1994);",
      "journal-title": "Genes Dev."
    },
    {
      "key": "e_1_3_1_2_4",
      "doi-asserted-by": "crossref",
      "first-page": "223",
      "DOI": "10.1038/370223a0",
      "volume": "370",
      "author": "Kwok R. P.",
      "year": "1994",
      "unstructured": "Kwok R. P., et al., Nature 370, 223 (1994).",
      "journal-title": "Nature"
    },
    {
      "key": "e_1_3_1_3_2",
      "doi-asserted-by": "crossref",
      "first-page": "403",
      "DOI": "10.1016/S0092-8674(00)81118-6",
      "volume": "85",
      "author": "Kamei Y.",
      "year": "1996",
      "unstructured": "Kamei Y., et al., Cell 85, 403 (1996).",
      "journal-title": "Cell"
    },
    {
      "key": "e_1_3_1_4_2",
      "doi-asserted-by": "crossref",
      "first-page": "99",
      "DOI": "10.1038/383099a0",
      "volume": "383",
      "author": "Chakravarti D.",
      "year": "1996",
      "unstructured": "Chakravarti D., et al., Nature 383, 99 (1996);",
      "journal-title": "Nature"
    },
    {
      "key": "e_1_3_1_4_3",
      "doi-asserted-by": "crossref",
      "first-page": "11540",
      "DOI": "10.1073/pnas.93.21.11540",
      "volume": "93",
      "author": "Hanstein B.",
      "year": "1996",
      "unstructured": "Hanstein B., et al., Proc. Natl. Acad. Sci. U.S.A. 93, 11540 (1996);",
      "journal-title": "Proc. Natl. Acad. Sci. U.S.A."
    },
    {
      "key": "e_1_3_1_4_4",
      "unstructured": "; T. P. Yao G. Ku N. Zhou R. Scully D. M. Livingston ibid. p. 10626."
    },
    {
      "key": "e_1_3_1_5_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.8197458"
    },
    {
      "key": "e_1_3_1_5_3",
      "doi-asserted-by": "crossref",
      "first-page": "10009",
      "DOI": "10.1073/pnas.91.21.10009",
      "volume": "91",
      "author": "Cavailles V.",
      "year": "1994",
      "unstructured": "Cavailles V., Dauvois S., Danielian P. S., Parker M. G., Proc. Natl. Acad. Sci. U.S.A. 91, 10009 (1994).",
      "journal-title": "Proc. Natl. Acad. Sci. U.S.A."
    },
    {
      "key": "e_1_3_1_6_2",
      "doi-asserted-by": "crossref",
      "unstructured": "S. A. Onate S. Y. Tsai M.-J. Tsai B. W. O'Malley Science 270 1354 (1995).",
      "DOI": "10.1126/science.270.5240.1354"
    },
    {
      "key": "e_1_3_1_7_2",
      "doi-asserted-by": "crossref",
      "first-page": "3667",
      "DOI": "10.1002/j.1460-2075.1996.tb00736.x",
      "volume": "15",
      "author": "Voegel J. J.",
      "year": "1996",
      "unstructured": "Voegel J. J., Heine M. J., Zechel C., Chambon P., Gronemeyer H., EMBO J. 15, 3667 (1996);",
      "journal-title": "EMBO J."
    },
    {
      "key": "e_1_3_1_7_3",
      "doi-asserted-by": "crossref",
      "first-page": "2735",
      "DOI": "10.1128/MCB.17.5.2735",
      "volume": "17",
      "author": "Hong H.",
      "year": "1997",
      "unstructured": "Hong H., Kohli K., Garabedian M. J., Stallcup M. R., Mol. Cell. Biol. 17, 2735 (1997).",
      "journal-title": "Mol. Cell. Biol."
    },
    {
      "key": "e_1_3_1_8_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/42652"
    },
    {
      "key": "e_1_3_1_9_2",
      "unstructured": "A human homolog of mouse p/CIP gene has been also described ["
    },
    {
      "key": "e_1_3_1_9_3",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0092-8674(00)80516-4"
    },
    {
      "key": "e_1_3_1_9_4",
      "doi-asserted-by": "crossref",
      "first-page": "8479",
      "DOI": "10.1073/pnas.94.16.8479",
      "volume": "94",
      "author": "Li H.",
      "year": "1997",
      "unstructured": "Li H., Gomes P. J., Chen J. D., Proc. Natl. Acad. Sci. U.S.A. 94, 8479 (1997);",
      "journal-title": "Proc. Natl. Acad. Sci. U.S.A."
    },
    {
      "key": "e_1_3_1_9_5",
      "unstructured": "; O. M. Sobulo et al. ibid. p. 8732]."
    },
    {
      "key": "e_1_3_1_10_2",
      "doi-asserted-by": "crossref",
      "first-page": "319",
      "DOI": "10.1038/382319a0",
      "volume": "382",
      "author": "Yang X. J.",
      "year": "1996",
      "unstructured": "Yang X. J., Ogryzko V. V., Nishikawa J., Howard B. H., Nakatani Y., Nature 382, 319 (1996).",
      "journal-title": "Nature"
    },
    {
      "key": "e_1_3_1_11_2",
      "doi-asserted-by": "crossref",
      "first-page": "4807",
      "DOI": "10.1002/j.1460-2075.1994.tb06806.x",
      "volume": "13",
      "author": "Marcus G. A.",
      "year": "1994",
      "unstructured": "Marcus G. A., Silverman N., Berger S. L., Horiuchi J., Guarente L., EMBO J. 13, 4807 (1994);",
      "journal-title": "EMBO J."
    },
    {
      "key": "e_1_3_1_11_3",
      "doi-asserted-by": "crossref",
      "first-page": "593",
      "DOI": "10.1128/MCB.16.2.593",
      "volume": "16",
      "author": "Candau R.",
      "year": "1996",
      "unstructured": "Candau R., et al., Mol. Cell. Biol. 16, 593 (1996);",
      "journal-title": "Mol. Cell. Biol."
    },
    {
      "key": "e_1_3_1_11_4",
      "doi-asserted-by": "crossref",
      "first-page": "5237",
      "DOI": "10.1074/jbc.271.9.5237",
      "volume": "271",
      "author": "Candau R.",
      "year": "1996",
      "unstructured": "Candau R., Berger S. L., J. Biol. Chem. 271, 5237 (1996);",
      "journal-title": "J. Biol. Chem."
    },
    {
      "key": "e_1_3_1_11_5",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0092-8674(00)81063-6"
    },
    {
      "key": "e_1_3_1_11_6",
      "doi-asserted-by": "crossref",
      "first-page": "555",
      "DOI": "10.1093/emboj/16.3.555",
      "volume": "16",
      "author": "Candau R.",
      "year": "1997",
      "unstructured": "Candau R., Zhou J. X., Allis C. D., Berger S. L., EMBO J. 16, 555 (1997).",
      "journal-title": "EMBO J."
    },
    {
      "key": "e_1_3_1_12_2",
      "doi-asserted-by": "crossref",
      "first-page": "953",
      "DOI": "10.1016/S0092-8674(00)82001-2",
      "volume": "87",
      "author": "Ogryzko V. V.",
      "year": "1996",
      "unstructured": "Ogryzko V. V., Schiltz R. L., Russanova V., Howard B. H., Nakatani Y., Cell 87, 953 (1996);",
      "journal-title": "Cell"
    },
    {
      "key": "e_1_3_1_12_3",
      "doi-asserted-by": "crossref",
      "first-page": "641",
      "DOI": "10.1038/384641a0",
      "volume": "384",
      "author": "Bannister A. J.",
      "year": "1996",
      "unstructured": "Bannister A. J., Kouzarides T., Nature 384, 641 (1996).",
      "journal-title": "Nature"
    },
    {
      "key": "e_1_3_1_13_2",
      "doi-asserted-by": "crossref",
      "first-page": "194",
      "DOI": "10.1038/38304",
      "volume": "389",
      "author": "Spencer T. E.",
      "year": "1997",
      "unstructured": "Spencer T. E., et al., Nature 389, 194 (1997).",
      "journal-title": "Nature"
    },
    {
      "key": "e_1_3_1_14_2",
      "first-page": "733",
      "volume": "387",
      "author": "Heery D. M.",
      "year": "1997",
      "unstructured": "Heery D. M., Kalkhoven E., Hoare S., Parker M., ibid. 387, 733 (1997).",
      "journal-title": "ibid."
    },
    {
      "key": "e_1_3_1_15_2",
      "doi-asserted-by": "crossref",
      "first-page": "697",
      "DOI": "10.1016/0092-8674(89)90016-0",
      "volume": "59",
      "author": "Glass C. K.",
      "year": "1989",
      "unstructured": "Glass C. K., Lipkin S. M., Devary O. V., Rosenfeld M. G., Cell 59, 697 (1989);",
      "journal-title": "Cell"
    },
    {
      "key": "e_1_3_1_15_3",
      "doi-asserted-by": "crossref",
      "first-page": "451",
      "DOI": "10.1038/377451a0",
      "volume": "377",
      "author": "Kurokawa R.",
      "year": "1995",
      "unstructured": "Kurokawa R., et al., Nature 377, 451 (1995).",
      "journal-title": "Nature"
    },
    {
      "key": "e_1_3_1_16_2",
      "doi-asserted-by": "crossref",
      "first-page": "397",
      "DOI": "10.1038/377397a0",
      "volume": "377",
      "author": "Horlein A. J.",
      "year": "1995",
      "unstructured": "Horlein A. J., et al., Nature 377, 397 (1995).",
      "journal-title": "Nature"
    },
    {
      "key": "e_1_3_1_17_2",
      "first-page": "49",
      "volume": "387",
      "author": "Alland L.",
      "year": "1997",
      "unstructured": "Alland L., et al., ibid. 387, 49 (1997);",
      "journal-title": "ibid."
    },
    {
      "key": "e_1_3_1_17_3",
      "doi-asserted-by": "crossref",
      "first-page": "341",
      "DOI": "10.1016/S0092-8674(00)80214-7",
      "volume": "89",
      "author": "Hassig C. A.",
      "year": "1997",
      "unstructured": "Hassig C. A., Fleischer T. C., Billin A. N., Schreiber S. L., Ayer D. E., Cell 89, 341 (1997);",
      "journal-title": "Cell"
    },
    {
      "key": "e_1_3_1_17_4",
      "doi-asserted-by": "crossref",
      "first-page": "43",
      "DOI": "10.1038/387043a0",
      "volume": "387",
      "author": "Heinzel T.",
      "year": "1997",
      "unstructured": "Heinzel T., et al., Nature 387, 43 (1997);",
      "journal-title": "Nature"
    },
    {
      "key": "e_1_3_1_17_5",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0092-8674(00)80217-2"
    },
    {
      "key": "e_1_3_1_17_6",
      "unstructured": "; C. D. Laherty et al. ibid. p. 349; L. Nagy et al. ibid. p. 373; Y. Zhang R. Iratni H. Erdjument-Bromage P. Tempst D. Reinberg ibid. p. 357."
    },
    {
      "key": "e_1_3_1_18_2",
      "doi-asserted-by": "crossref",
      "first-page": "1025",
      "DOI": "10.1002/j.1460-2075.1992.tb05141.x",
      "volume": "11",
      "author": "Danielian P. S.",
      "year": "1992",
      "unstructured": "Danielian P. S., White R., Lees J. A., Parker M. G., EMBO J. 11, 1025 (1992);",
      "journal-title": "EMBO J."
    },
    {
      "key": "e_1_3_1_18_3",
      "first-page": "5370",
      "volume": "13",
      "author": "Durand B.",
      "year": "1994",
      "unstructured": "Durand B., et al., ibid. 13, 5370 (1994).",
      "journal-title": "ibid."
    },
    {
      "key": "e_1_3_1_19_2",
      "doi-asserted-by": "crossref",
      "first-page": "251",
      "DOI": "10.1016/0092-8674(92)90100-Q",
      "volume": "70",
      "author": "Berger S. L.",
      "year": "1992",
      "unstructured": "Berger S. L., et al., Cell 70, 251 (1992);",
      "journal-title": "Cell"
    },
    {
      "key": "e_1_3_1_19_3",
      "doi-asserted-by": "crossref",
      "first-page": "4145",
      "DOI": "10.1002/j.1460-2075.1992.tb05507.x",
      "volume": "11",
      "author": "Georgakopoulos T.",
      "year": "1992",
      "unstructured": "Georgakopoulos T., Thireos G., EMBO J. 11, 4145 (1992).",
      "journal-title": "EMBO J."
    },
    {
      "key": "e_1_3_1_20_2",
      "doi-asserted-by": "crossref",
      "first-page": "28138",
      "DOI": "10.1074/jbc.271.45.28138",
      "volume": "271",
      "author": "Swope D. L.",
      "year": "1996",
      "unstructured": "Swope D. L., Mueller C. L., Chrivia J. C., J. Biol. Chem. 271, 28138 (1996);",
      "journal-title": "J. Biol. Chem."
    },
    {
      "key": "e_1_3_1_20_3",
      "unstructured": "; S. Bisotto S. Minorgan R. P. Rehfuss ibid. p. 17746."
    },
    {
      "key": "e_1_3_1_21_2",
      "unstructured": "E. Korzus D. W. Rose M. G. Rosenfeld data not shown."
    },
    {
      "key": "e_1_3_1_22_2",
      "doi-asserted-by": "crossref",
      "first-page": "154",
      "DOI": "10.1016/S0968-0004(97)01034-7",
      "volume": "22",
      "author": "Neuwald A. F.",
      "year": "1997",
      "unstructured": "Neuwald A. F., Landsman D., Trends Biochem. Sci. 22, 154 (1997).",
      "journal-title": "Trends Biochem. Sci."
    },
    {
      "key": "e_1_3_1_23_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.279.5351.700"
    },
    {
      "key": "e_1_3_1_24_2",
      "doi-asserted-by": "crossref",
      "first-page": "15092",
      "DOI": "10.1073/pnas.93.26.15092",
      "volume": "93",
      "author": "Zhang J. J.",
      "year": "1996",
      "unstructured": "Zhang J. J., et al., Proc. Natl. Acad. Sci. U.S.A. 93, 15092 (1996);",
      "journal-title": "Proc. Natl. Acad. Sci. U.S.A."
    },
    {
      "key": "e_1_3_1_24_3",
      "doi-asserted-by": "crossref",
      "first-page": "344",
      "DOI": "10.1038/383344a0",
      "volume": "383",
      "author": "Bhattacharya S.",
      "year": "1996",
      "unstructured": "Bhattacharya S., et al., Nature 383, 344 (1996);",
      "journal-title": "Nature"
    },
    {
      "key": "e_1_3_1_24_4",
      "doi-asserted-by": "crossref",
      "first-page": "1074",
      "DOI": "10.1073/pnas.94.4.1074",
      "volume": "94",
      "author": "Horvai A. E.",
      "year": "1997",
      "unstructured": "Horvai A. E., et al., Proc. Natl. Acad. Sci. U.S.A. 94, 1074 (1997).",
      "journal-title": "Proc. Natl. Acad. Sci. U.S.A."
    },
    {
      "key": "e_1_3_1_25_2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.277.5332.1630"
    },
    {
      "key": "e_1_3_1_26_2",
      "unstructured": "Whole-cell extracts were prepared by lysing the cells in NET-N buffer containing 50 mM tris (pH 7.6) 5 mM EDTA 0.3 M NaCl 1 mM dithiothreitol 0.1% NP-40 and protease inhibitors (2 mM phenylmethylsulfonyl fluoride and 10 \u03bcg/ml each of leupeptin pepstatin and aprotinin). The mixture was then centrifuged at 30 000 g for 1 hour at 4\u00b0C and the supernatant was stored at \u201380\u00b0C until use. For coimmunoprecipitation assays 1 mg of cell extract was incubated in the presence of 2 \u03bcg of anti-p/CIP anti-Flag or anti-RAR IgG for 2 hours at 4\u00b0C. The immune complexes were then precipitated with protein A\u2013Sepharose. Protein complexes were resolved by SDS\u2013polyacrylamide gel electrophoresis (PAGE) ["
    },
    {
      "key": "e_1_3_1_26_3",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/227680a0"
    },
    {
      "key": "e_1_3_1_26_4",
      "unstructured": "] protein immunoblotted and probed using 1 \u03bcg/ml of anti-Flag anti-p/CIP or an anti-hemagglutinin (anti-HA)."
    },
    {
      "key": "e_1_3_1_27_2",
      "unstructured": "The yeast strain EGY 48 the LexA-b galactosidase reporter construct (PSH 18-34) and the B42 parental vectors (PEG 202 and PJG 4-5) were all previously described ["
    },
    {
      "key": "e_1_3_1_27_3",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(93)90498-F"
    },
    {
      "key": "e_1_3_1_27_4",
      "unstructured": "]. Various p/CAF fragments or fragments of other coactivators were obtained by polymerase chain reaction or restriction digestion and subcloned into PEG 202 bait vector or PJG 4-5 prey vectors respectively. EGY 48 cells were transformed with the lac Z reporter plasmid pSH 18-34 with the appropriate bait and prey vectors and were plated out on Ura \u2013 His \u2013 Trp \u2013 medium containing 2% galactose. Isolated yeast colonies were then allowed to grow in the same liquid medium and were subsequently assayed for \u03b2-galactosidase as previously described [F. M. Ausubel et al. Current Protocols in Molecular Biology (Wiley New York 1995)]. All constructs were sequenced and functionally tested in yeast two-hybrid assays showing interaction with other proteins as previously described (2 7). In the case of p/CAF constructs each fragment interacted positively with at least one other coactivator except the fragment with amino acids 86 through 518."
    },
    {
      "key": "e_1_3_1_28_2",
      "unstructured": "Insulin-responsive Rat-1 fibroblasts were seeded on acid-washed glass cover slips at subconfluent density and grown in D-MEM/F-12 (Life Technologies) medium supplemented with 10% fetal bovine serum gentamicin and methotrexate. Before the injection the cells were rendered quiescent by incubation in serum-free medium for 24 to 36 hours. Plasmids were injected into the nuclei of cells at a final concentration of 100 \u03bcg/ml. Immunoglobulin G specific for p/CAF was prepared from guinea pig serum raised against a bacterially expressed fragment (amino acids 466 to 832) of p/CAF; this IgG recognized the single band of p/CAF in protein immunoblot analysis. Either preimmune IgG or the appropriate specific antibodies directed against p/CAF p/CIP SRC-1 or CBP were coinjected and allowed the unambiguous identification of the injected cells (7). Preimmune controls were included in all experiments. Microinjections were carried out using an Eppendorf semiautomated microinjection system mounted on an inverted Zeiss microscope. Approximately 1 hour after injection the cells were stimulated where indicated with the appropriate ligand. In the case of rescue experiments the cells were stimulated with ligand 6 hours after injection to allow protein expression. After overnight incubation the cells were fixed and then stained to detect injected IgG and \u03b2-galactosidase expression ["
    },
    {
      "key": "e_1_3_1_28_3",
      "doi-asserted-by": "crossref",
      "first-page": "1405",
      "DOI": "10.1083/jcb.119.6.1405",
      "volume": "119",
      "author": "Rose D. W.",
      "year": "1992",
      "unstructured": "Rose D. W., et al., J. Cell Biol. 119, 1405 (1992);",
      "journal-title": "J. Cell Biol."
    },
    {
      "key": "e_1_3_1_28_4",
      "unstructured": "; (2)]. Injected cells were identified by staining with tetramethylrhodamine-conjugated donkey anti-rabbit IgG. All experimental results are expressed as the mean \u00b1 SEM of at least three experiments in which at least 1000 cells were injected."
    },
    {
      "key": "e_1_3_1_29_2",
      "unstructured": "GST-RAR and GST-CBP fragments were generated as described (2). We incubated 25 \u03bcl of GST-Sepharose beads containing 3 to 6 \u03bcg of the GST recombinant proteins with 5 \u00d7 10 5 cpm of 35 S-labeled p/CAF proteins generated by in vitro transcription and translation for 2 hours at 4\u00b0C. The complexes were washed five times with NET-N buffer resolved by SDS-PAGE and fluorographed."
    },
    {
      "key": "e_1_3_1_30_2",
      "unstructured": "Mutations in p/CAF and CBP were introduced by site-directed mutagenesis using the Quick-Change mutagenesis system (Stratagene) according to the manufacturer's instructions. Double-stranded oligonucleotides were designed such that the wild-type sequence corresponding to amino acids Tyr 616 /Phe 617 (acetyl-CoA\u2013binding site) in p/CAF cDNA were substituted with alanines in order to generate a mutant of p/CAF lacking HAT activity (pCMV\u2013 p/CAF HAT\u2013 ). A similar strategy was used to obtain mutants of CBP. Mutants of p/CAF and CBP were expressed in bacteria and baculovirus respectively and tested for HAT activity in solution using histones as substrates ["
    },
    {
      "key": "e_1_3_1_30_3",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.92.14.6364"
    },
    {
      "key": "e_1_3_1_31_2",
      "unstructured": "We thank R. Heyman for use of TTNPB and LG629 S. L. Berger for discussion and providing hGCN5 expression vector and Y. Nakatani and X. J. Yang for providing a tagged p/CAF expression vector. We also thank C. Nelson for experimental assistance L.-M. Phillips for excellent technical assistance and P. Myer for expertise in figure preparation. E.K. is supported by a U.S. Army Medical Research Program Award J.T. by the National Cancer Institute of Canada E.M.M. by an NIH Postdoctoral Fellowship D.W.R. by an American Diabetes Association Career Development Award and L.X. by an American Heart Association Predoctoral Fellowship. Supported by grants from NIH to C.K.G. and M.G.R."
    }
  ],
  "container-title": "Science",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://www.science.org/doi/pdf/10.1126/science.279.5351.703",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2024,
        1,
        13
      ]
    ],
    "date-time": "2024-01-13T05:58:03Z",
    "timestamp": 1705125483000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "https://www.science.org/doi/10.1126/science.279.5351.703"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        1998,
        1,
        30
      ]
    ]
  },
  "references-count": 61,
  "journal-issue": {
    "issue": "5351",
    "published-print": {
      "date-parts": [
        [
          1998,
          1,
          30
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1126/science.279.5351.703"
  ],
  "URL": "http://dx.doi.org/10.1126/science.279.5351.703",
  "relation": {},
  "ISSN": [
    "0036-8075",
    "1095-9203"
  ],
  "subject": [],
  "container-title-short": "Science",
  "published": {
    "date-parts": [
      [
        1998,
        1,
        30
      ]
    ]
  }
}