DOI: 10.1126/science.280.5361.298. Coupling Termination of Transcription to Messenger RNA Maturation in Yeast

Coupling Termination of Transcription to Messenger RNA Maturation in Yeast

10.1126/science.280.5361.298

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

The direct association between messenger RNA (mRNA) 3′-end processing and the termination of transcription was established for the CYC1 gene of Saccharomyces cerevisiae . The mutation of factors involved in the initial cleavage of the primary transcript at the poly(A) site ( RNA14 , RNA15 , and PCF11 ) disrupted transcription termination at the 3′ end of the CYC1 gene. In contrast, the mutation of factors involved in the subsequent polyadenylation step ( PAP1 , FIP1 , and YTH1 ) had little effect. Thus, cleavage factors link transcription termination of RNA polymerase II with pre-mRNA 3′-end processing.

Bibliography

Birse, C. E., Minvielle-Sebastia, L., Lee, B. A., Keller, W., & Proudfoot, N. J. (1998). Coupling Termination of Transcription to Messenger RNA Maturation in Yeast. Science, 280(5361), 298â301.

Authors 5

Charles E. Birse (first)
Lionel Minvielle-Sebastia (additional)
Barbara A. Lee (additional)
Walter Keller (additional)
Nick J. Proudfoot (additional)

References 22 Referenced 204

E. Whitelaw, N. J. Proudfoot, EMBO J. 5, 2915 (1986); J. Logan, E. Falck-Pedersen, J. E. Darnell, T. Shenk, Proc. Natl. Acad. Sci. U.S.A. 84, 8306 (1987); S. Connelly, J. L. Manley, Genes Dev. 2, 440 (1988); Lanoix J., Acheson N. H., EMBO J. 7, 2515 (1988). (10.1002/j.1460-2075.1988.tb03099.x) / EMBO J. by Lanoix J. (1988)
P. Russo, F. Sherman, Proc. Natl. Acad. Sci. U.S.A. 86, 8348 (1989); Russo P., Yeast 11, 447 (1995). (10.1002/yea.320110507) / Yeast by Russo P. (1995)
Birse C. E., Lee B. A., Hansen K., Proudfoot N. J., EMBO J.16, 3633 (1997). (10.1093/emboj/16.12.3633) / EMBO J. by Birse C. E. (1997)
J. L. Manley, Y. Takagaki, Science 274, 1481 (1996); Keller W., Minvielle-Sebastia L., Curr. Opin. Cell Biol. 9, 329 (1997). (10.1016/S0955-0674(97)80004-X) / Curr. Opin. Cell Biol. by Keller W. (1997)
Chen J., Moore C., Mol. Cell. Biol.12, 3470 (1992). / Mol. Cell. Biol. by Chen J. (1992)
L. Minvielle-Sebastia, P. J. Preker, W. Keller, Science 266, 1702 (1994); P. J. Preker, J. Lingner, L. Minvielle-Sebastia, W. Keller, Cell 81, 379 (1995); M. M. Kessler, J. Zhao, C. L. Moore, J. Biol. Chem. 271, 27167 (1996); N. Amrani, et al., Mol. Cell. Biol. 17, 1102 (1997); P. J. Preker, M. Ohnacker, L.Minvielle-Sebastia, W. Keller, EMBO J. 16, 4727 (1997); J. Zhao, M. M. Kessler, C. L. Moore, J. Biol. Chem. 272, 10831 (1997); M. M. Kessler, et al., Genes Dev. 11, 2545 (1997); D. Patel, J. S. Butler, Mol. Cell. Biol. 12, 3297 (1992); Barabino M. L., Hubner W., Jenny A., Minvielle-Sebastia L., Keller W., Genes Dev. 11, 1703 (1997). (10.1101/gad.11.13.1703) / Genes Dev. by Barabino M. L. (1997)
Guo Z., Sherman F., Trends Biochem. Sci.21, 477 (1996). (10.1016/S0968-0004(96)10057-8) / Trends Biochem. Sci. by Guo Z. (1996)
Yeast strains were cultured in selective media (YNB) supplemented with appropriate amino acids [C. Guthrie, G. R. Fink, Methods Enzymol. 194, 14 (1991)] with galactose (2%) as a carbon source. Transformation was performed as described [Gietz D., St. Jean A., Woods R. A., Schiestl R. H., Nucleic Acids Res. 20, 1425 (1992)]. Strains RY260 and SB7 were transformed with plasmid pUGCYC, which carries the URA3 (and TRP1) selectable marker. All other strains were transformed with pGCYC carryingTRP1. Conditional strains were grown at the permissive temperature (22° to 25°C) to an optical density at 600 nm of 0.1 to 0.2, briefly harvested by centrifugation (1 min), and resuspended in prewarmed media (37°C) before being cultured at the nonpermissive temperature for 5, 25, or 60 min. Formamide (2%) was added to media to restrict the function of the fip1-1 andyth1-1 alleles (6). (10.1093/nar/20.6.1425) / Nucleic Acids Res. by Gietz D. (1992)
To form pGCYC, we used PCR to synthesize the region extending from 33 to 1962 bp 3′ of the CYC1 open reading frame (ORF) start site, with genomic DNA as a template. This fragment was cloned into the unique Bgl II site of pRS424.KV, which contains the GAL1/10 promoter region (C. Goding, unpublished data) cloned into the shuttle vector pRS424 [Christianson T. W., Sikorski R. S., Dante M., Shero J. H., Hieter P., Gene 110, 119 (1992)]. A 1480-bp fragment containing the URA3 gene (spanning the region from 500 bp upstream of the URA3 ORF start site to 176 bp downstream of the translation stop site) was next ligated into the Bam HI site upstream of the GAL1/10 promoter to generate pUGCYC. pGcyc1-512 was constructed by PCR and contains the originally identified poly(A) site deletion mutation,cyc1-512 (14). (10.1016/0378-1119(92)90454-W) / Gene by Christianson T. W. (1992)
Heidmann S., Obermaier B., Vogel K., Domdey H., Mol. Cell. Biol.12, 4215 (1992). / Mol. Cell. Biol. by Heidmann S. (1992)
C. E. Birse B. A. Lee N. J. Proudfoot L. Minvielle-Sebastia W. Keller unpublished data.
Barry K., Stiles J. I., Pietras D. F., Melnick I., Sherman F., Mol. Cell. Biol.7, 632 (1987). / Mol. Cell. Biol. by Barry K. (1987)
M. Nonet C. Scafe J. Sexton R. Young ibid. p. 1602.
Zaret K. S., Sherman F., Cell28, 563 (1982). (10.1016/0092-8674(82)90211-2) / Cell by Zaret K. S. (1982)
Edwalds-Gilbert G., Prescott J., Falck-Pedersen E., Mol. Cell. Biol.13, 3472 (1993). / Mol. Cell. Biol. by Edwalds-Gilbert G. (1993)
Minvielle-Sebastia L., Winsor B., Bonneaud N., Lacroute F., ibid11, 3075 (1991). / ibid by Minvielle-Sebastia L. (1991)
N. Amrani, M. Minet, M. Le Gouar, F. Lacroute, F. Wyers,ibid. 17, 3674 (1997); Minvielle-Sebastia L., Preker P. J., Wiederkehr T., Strahm Y., Keller W., Proc. Natl. Acad. Sci. U.S.A. 94, 7897 (1997). (10.1073/pnas.94.15.7897) / Proc. Natl. Acad. Sci. U.S.A. by Minvielle-Sebastia L. (1997)
Sachs A., Deardorff J. A., Cell70, 961 (1992). (10.1016/0092-8674(92)90246-9) / Cell by Sachs A. (1992)
10.1038/38763
The six contiguous M13 clones spanning the 3′ end of the CYC1 gene were constructed from PCR products with pGCYC as a template. The 172-bp GAL probe used to detect transcripts upstream of the GAL promoter was generated with pGCYC as a PCR template. The tRNA probe was generated with genomic DNA as a PCR template. This 225-bp fragment spans the SUP11 gene coding for Tyr-tRNA located on chromosome 6. The actin ACT1 probe was generated as a 567-bp fragment encompassing the region 277 to 844 bp 3′of the ACT1 ORF start site. All fragments were cloned into M13mp19 (RF) restricted with Hinc II. The M13 control probe has no insert.
The wild-type Rna15p and temperature-sensitive Rna15-1p proteins were synthesized in vitro in rabbit reticulocyte lysates with [35S]methionine. After incubation for 10 min at 45°C, they were assayed for their ability to bind to agarose-bound poly(U) for 15 min, as described [Swanson M. S., Dreyfuss G., Mol. Cell. Biol. 8, 2237 (1988)]. After extensive washes, the bound material was released by boiling with SDS– polyacrylamide gel electrophoresis (PAGE) loading buffer and loaded on 10% denaturing protein gels. After electrophoresis, the gels were fixed, dried, and subjected to autoradiography. / Mol. Cell. Biol. by Swanson M. S. (1988)
We thank S. Barabino S. Butler F. Lacroute A. Sachs F. Sherman and R. Young for various yeast strains P. J. Preker for the pap1 mutants and J. Mellor for plasmids. We thank D. Pritlove and members of the N.J.P. lab for critical reading of the manuscript and for their help and advice. Supported by a Wellcome Programme Grant (032773) to N.J.P. and grants by the Kantons of Basel the Swiss National Science Foundation and the European Union (through the Bundesamt für Bildung und Wissenschaft Bern) to W.K.

Dates

Type	When
Created	23 years ago (July 27, 2002, 5:49 a.m.)
Deposited	1 year, 7 months ago (Jan. 13, 2024, 12:12 a.m.)
Indexed	2 weeks, 6 days ago (Aug. 3, 2025, 12:35 a.m.)
Issued	27 years, 4 months ago (April 10, 1998)
Published	27 years, 4 months ago (April 10, 1998)
Published Print	27 years, 4 months ago (April 10, 1998)

Funders 0

None

BibTeX

@article{Birse_1998, title={Coupling Termination of Transcription to Messenger RNA Maturation in Yeast}, volume={280}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.280.5361.298}, DOI={10.1126/science.280.5361.298}, number={5361}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Birse, Charles E. and Minvielle-Sebastia, Lionel and Lee, Barbara A. and Keller, Walter and Proudfoot, Nick J.}, year={1998}, month=apr, pages={298–301} }

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  "abstract": "<jats:p>\n            The direct association between messenger RNA (mRNA) 3\u2032-end processing and the termination of transcription was established for the\n            <jats:italic>CYC1</jats:italic>\n            gene of\n            <jats:italic>Saccharomyces cerevisiae</jats:italic>\n            . The mutation of factors involved in the initial cleavage of the primary transcript at the poly(A) site (\n            <jats:italic>RNA14</jats:italic>\n            ,\n            <jats:italic>RNA15</jats:italic>\n            , and\n            <jats:italic>PCF11</jats:italic>\n            ) disrupted transcription termination at the 3\u2032 end of the\n            <jats:italic>CYC1</jats:italic>\n            gene. In contrast, the mutation of factors involved in the subsequent polyadenylation step (\n            <jats:italic>PAP1</jats:italic>\n            ,\n            <jats:italic>FIP1</jats:italic>\n            , and\n            <jats:italic>YTH1</jats:italic>\n            ) had little effect. Thus, cleavage factors link transcription termination of RNA polymerase II with pre-mRNA 3\u2032-end processing.\n          </jats:p>",
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    },
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      "key": "e_1_3_1_22_2",
      "first-page": "2237",
      "volume": "8",
      "author": "Swanson M. S.",
      "year": "1988",
      "unstructured": "The wild-type Rna15p and temperature-sensitive Rna15-1p proteins were synthesized in vitro in rabbit reticulocyte lysates with [35S]methionine. After incubation for 10 min at 45\u00b0C, they were assayed for their ability to bind to agarose-bound poly(U) for 15 min, as described [Swanson M. S., Dreyfuss G., Mol. Cell. Biol. 8, 2237 (1988)]. After extensive washes, the bound material was released by boiling with SDS\u2013 polyacrylamide gel electrophoresis (PAGE) loading buffer and loaded on 10% denaturing protein gels. After electrophoresis, the gels were fixed, dried, and subjected to autoradiography.",
      "journal-title": "Mol. Cell. Biol."
    },
    {
      "key": "e_1_3_1_23_2",
      "unstructured": "We thank S. Barabino S. Butler F. Lacroute A. Sachs F. Sherman and R. Young for various yeast strains P. J. Preker for the pap1 mutants and J. Mellor for plasmids. We thank D. Pritlove and members of the N.J.P. lab for critical reading of the manuscript and for their help and advice. Supported by a Wellcome Programme Grant (032773) to N.J.P. and grants by the Kantons of Basel the Swiss National Science Foundation and the European Union (through the Bundesamt f\u00fcr Bildung und Wissenschaft Bern) to W.K."
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