DOI: 10.1101/gad.8.24.3008. Dynamic association of proteins with the pre-mRNA branch region.

Dynamic association of proteins with the pre-mRNA branch region.

10.1101/gad.8.24.3008

Crossref journal-article

Cold Spring Harbor Laboratory

Genes & Development (246)

Abstract

The association of proteins with the branch site region during pre-mRNA splicing was probed using a novel methodology to site-specifically modify the pre-mRNA with the photo-reagent benzophenone. Three sets of proteins were distinguished by the kinetics of their associations with pre-mRNAs, by their association with discrete splicing complexes, and by their differing factor requirements. An early U1 snRNP-dependent cross-link of the branch region to a p80 species was followed by cross-links to p14, p35, and p150 polypeptides associated with the U2 snRNP-pre-mRNA complex. Concomitant with formation of the spliceosome, a rearrangement of protein factors about the branch region occurred, in which the p35 and p150 cross-links were replaced by p220 and p70 species. These results establish that the branch region is recognized in a dynamic fashion by multiple distinct proteins during the course of spliceosomal assembly.

Bibliography

MacMillan, A. M., Query, C. C., Allerson, C. R., Chen, S., Verdine, G. L., & Sharp, P. A. (1994). Dynamic association of proteins with the pre-mRNA branch region. Genes & Development, 8(24), 3008â3020.

Authors 6

A M MacMillan (first)
C C Query (additional)
C R Allerson (additional)
S Chen (additional)
G L Verdine (additional)
P A Sharp (additional)

References 52 Referenced 104

10.1038/342819a0
10.1016/0092-8674(90)90162-8
10.1073/pnas.81.12.3607
10.1073/pnas.90.17.8229
{'key': '2021111418193757000_8.24.3008.5', 'first-page': '307', 'article-title': 'Small nuclear ribonucleoprotein (RNP) U2 contains numerous additional proteins and has a bipartite RNP structure under splicing conditions.', 'volume': '13', 'year': '1993', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Small nuclear ribonucleoprotein (RNP) U2 contains numerous additional proteins and has a bipartite RNP structure under splicing conditions. (1993)
Blackburn, P. and S. Moore. 1982. Pancreatic ribonuclease. In The enzymes (ed. P.D. Boyer), pp. 317â433. Academic Press, New York. (10.1016/S1874-6047(08)60284-X)
Blencowe, B.J. and S.M.L. Barabino. 1994. Antisense affinity depletion of RNP particles: Application to spliceosomal sn-RNPs. In Methods in molecular biology, Vol. XX: In vitro transcription and translation protocols (ed. M.J. Tymms). Humana Press, Totowa, NJ (in press).
10.1083/jcb.127.3.593
10.1126/science.8211112
10.1016/S0021-9258(19)85380-2 / J. Biol. Chem. / Separation of splicing factor SF3 into two components and purification of SF3a activity. (1993)
10.1101/gad.8.16.1974
{'key': '2021111418193757000_8.24.3008.12', 'first-page': '1866', 'article-title': 'Complementation by SR proteins of pre-mRNA splicing reactions depleted of U1 snRNP.', 'volume': '262', 'year': '1994', 'journal-title': 'Science'} / Science / Complementation by SR proteins of pre-mRNA splicing reactions depleted of U1 snRNP. (1994)
10.1093/nar/11.5.1475
10.1038/335375a0
10.1021/bi00185a001
10.1101/gad.3.12a.1874
10.1073/pnas.87.8.3082
10.1016/0092-8674(84)90372-6
10.1126/science.1853200
10.1016/S0960-9822(00)00061-0
10.1073/pnas.89.12.5482
10.1016/0092-8674(86)90066-8
10.1016/0092-8674(87)90614-3
10.1016/0167-4781(93)90122-T / Biochim. Biophys. Acta / Non-snRNP protein splicing factors. (1993)
10.1073/pnas.76.11.5495
10.1016/0092-8674(92)90556-R
10.1101/gad.8.9.1071
10.1016/0092-8674(92)90557-S
10.1101/gad.5.12b.2534
10.1126/science.1589782
Moore, M.J., C.C. Query, and P.A Sharp. 1993. Splicing of precursors to mRNA by the spliceosome. In The RNA world (ed. R.F. Gesteland and J.F. Atkins), pp. 303â357. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.
10.1021/bi00169a019
10.1016/0092-8674(94)90563-0
10.1016/0092-8674(87)90564-2
10.1101/gad.5.7.1224
Query, C.C. 1990. âAnalysis of RNA-protein interactions by identification of an RNA recognition motif within the U1 snRNA 70K autoantigen and definition of a protein-binding domain of U1 RNA,â Ph.D. thesis, Duke University, Durham, NC.
10.1101/gad.8.5.587
10.1101/gad.2.10.1268
10.1126/science.2973660
10.1016/0092-8674(88)90509-0
10.1016/0092-8674(89)90296-1
10.1002/j.1460-2075.1991.tb08062.x / EMBO J. / The yeast branchpoint sequence is not required for the formation of a stable U1 snRNP pre-mRNA complex and is recognized in the absence of U2 snRNA. (1991)
{'key': '2021111418193757000_8.24.3008.43', 'first-page': '2994', 'article-title': 'Direct interactions between pre-mRNA and six U2 small nuclear ribonucleoproteins during spliceosome assembly.', 'volume': '14', 'year': '1994', 'journal-title': 'Mol. Cell. Biol.'} / Mol. Cell. Biol. / Direct interactions between pre-mRNA and six U2 small nuclear ribonucleoproteins during spliceosome assembly. (1994)
Takahashi, K. and S. Moore. 1982. Ribonuclease T1. In The enzymes (ed. P.D. Boyer), pp. 435â468. Academic Press, New York. (10.1016/S1874-6047(08)60285-1)
10.1073/pnas.91.2.594
10.1016/S0021-9258(19)57239-8 / J. Biol. Chem. / Autoantibodies reactive with U1 RNA and initiator methionine transfer RNA. (1986)
10.1101/gad.3.10.1553
10.1101/gad.6.12b.2542
10.1002/j.1460-2075.1991.tb07937.x / EMBO J. / Biochemical characterization of U2 snRNP auxiliary factor: An essential pre-mRNA splicing factor with a novel intranuclear distribution. (1991)
10.1038/355609a0
10.1101/gad.3.10.1545
10.1073/pnas.86.8.2752

Dates

Type	When
Created	18 years, 2 months ago (June 5, 2007, 5:15 p.m.)
Deposited	3 years, 9 months ago (Nov. 14, 2021, 9:28 p.m.)
Indexed	3 months, 2 weeks ago (May 7, 2025, 4 p.m.)
Issued	30 years, 8 months ago (Dec. 15, 1994)
Published	30 years, 8 months ago (Dec. 15, 1994)
Published Online	30 years, 8 months ago (Dec. 15, 1994)
Published Print	30 years, 8 months ago (Dec. 15, 1994)

Funders 0

None

BibTeX

@article{MacMillan_1994, title={Dynamic association of proteins with the pre-mRNA branch region.}, volume={8}, ISSN={1549-5477}, url={http://dx.doi.org/10.1101/gad.8.24.3008}, DOI={10.1101/gad.8.24.3008}, number={24}, journal={Genes & Development}, publisher={Cold Spring Harbor Laboratory}, author={MacMillan, A M and Query, C C and Allerson, C R and Chen, S and Verdine, G L and Sharp, P A}, year={1994}, month=dec, pages={3008–3020} }

JSON

{
  "indexed": {
    "date-parts": [
      [
        2025,
        5,
        7
      ]
    ],
    "date-time": "2025-05-07T20:00:59Z",
    "timestamp": 1746648059412
  },
  "reference-count": 52,
  "publisher": "Cold Spring Harbor Laboratory",
  "issue": "24",
  "content-domain": {
    "domain": [],
    "crossmark-restriction": false
  },
  "published-print": {
    "date-parts": [
      [
        1994,
        12,
        15
      ]
    ]
  },
  "abstract": "<jats:p>The association of proteins with the branch site region during pre-mRNA splicing was probed using a novel methodology to site-specifically modify the pre-mRNA with the photo-reagent benzophenone. Three sets of proteins were distinguished by the kinetics of their associations with pre-mRNAs, by their association with discrete splicing complexes, and by their differing factor requirements. An early U1 snRNP-dependent cross-link of the branch region to a p80 species was followed by cross-links to p14, p35, and p150 polypeptides associated with the U2 snRNP-pre-mRNA complex. Concomitant with formation of the spliceosome, a rearrangement of protein factors about the branch region occurred, in which the p35 and p150 cross-links were replaced by p220 and p70 species. These results establish that the branch region is recognized in a dynamic fashion by multiple distinct proteins during the course of spliceosomal assembly.</jats:p>",
  "DOI": "10.1101/gad.8.24.3008",
  "type": "journal-article",
  "created": {
    "date-parts": [
      [
        2007,
        6,
        5
      ]
    ],
    "date-time": "2007-06-05T21:15:46Z",
    "timestamp": 1181078146000
  },
  "page": "3008-3020",
  "source": "Crossref",
  "is-referenced-by-count": 104,
  "title": "Dynamic association of proteins with the pre-mRNA branch region.",
  "prefix": "10.1101",
  "volume": "8",
  "author": [
    {
      "given": "A M",
      "family": "MacMillan",
      "sequence": "first",
      "affiliation": []
    },
    {
      "given": "C C",
      "family": "Query",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "C R",
      "family": "Allerson",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "S",
      "family": "Chen",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "G L",
      "family": "Verdine",
      "sequence": "additional",
      "affiliation": []
    },
    {
      "given": "P A",
      "family": "Sharp",
      "sequence": "additional",
      "affiliation": []
    }
  ],
  "member": "246",
  "published-online": {
    "date-parts": [
      [
        1994,
        12,
        15
      ]
    ]
  },
  "reference": [
    {
      "key": "2021111418193757000_8.24.3008.1",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/342819a0"
    },
    {
      "key": "2021111418193757000_8.24.3008.2",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(90)90162-8"
    },
    {
      "key": "2021111418193757000_8.24.3008.3",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.81.12.3607"
    },
    {
      "key": "2021111418193757000_8.24.3008.4",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.90.17.8229"
    },
    {
      "key": "2021111418193757000_8.24.3008.5",
      "first-page": "307",
      "article-title": "Small nuclear ribonucleoprotein (RNP) U2 contains numerous additional proteins and has a bipartite RNP structure under splicing conditions.",
      "volume": "13",
      "year": "1993",
      "journal-title": "Mol. Cell. Biol."
    },
    {
      "key": "2021111418193757000_8.24.3008.6",
      "doi-asserted-by": "crossref",
      "unstructured": "Blackburn, P. and S. Moore. 1982. Pancreatic ribonuclease. In The enzymes (ed. P.D. Boyer), pp. 317\u00e2\u0080\u0093433. Academic Press, New York.",
      "DOI": "10.1016/S1874-6047(08)60284-X"
    },
    {
      "key": "2021111418193757000_8.24.3008.7",
      "unstructured": "Blencowe, B.J. and S.M.L. Barabino. 1994. Antisense affinity depletion of RNP particles: Application to spliceosomal sn-RNPs. In Methods in molecular biology, Vol. XX: In vitro transcription and translation protocols (ed. M.J. Tymms). Humana Press, Totowa, NJ (in press)."
    },
    {
      "key": "2021111418193757000_8.24.3008.8",
      "doi-asserted-by": "publisher",
      "DOI": "10.1083/jcb.127.3.593"
    },
    {
      "key": "2021111418193757000_8.24.3008.9",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.8211112"
    },
    {
      "key": "2021111418193757000_8.24.3008.10",
      "doi-asserted-by": "crossref",
      "first-page": "17640",
      "DOI": "10.1016/S0021-9258(19)85380-2",
      "article-title": "Separation of splicing factor SF3 into two components and purification of SF3a activity.",
      "volume": "268",
      "year": "1993",
      "journal-title": "J. Biol. Chem."
    },
    {
      "key": "2021111418193757000_8.24.3008.11",
      "doi-asserted-by": "publisher",
      "DOI": "10.1101/gad.8.16.1974"
    },
    {
      "key": "2021111418193757000_8.24.3008.12",
      "first-page": "1866",
      "article-title": "Complementation by SR proteins of pre-mRNA splicing reactions depleted of U1 snRNP.",
      "volume": "262",
      "year": "1994",
      "journal-title": "Science"
    },
    {
      "key": "2021111418193757000_8.24.3008.13",
      "doi-asserted-by": "publisher",
      "DOI": "10.1093/nar/11.5.1475"
    },
    {
      "key": "2021111418193757000_8.24.3008.14",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/335375a0"
    },
    {
      "key": "2021111418193757000_8.24.3008.15",
      "doi-asserted-by": "publisher",
      "DOI": "10.1021/bi00185a001"
    },
    {
      "key": "2021111418193757000_8.24.3008.16",
      "doi-asserted-by": "publisher",
      "DOI": "10.1101/gad.3.12a.1874"
    },
    {
      "key": "2021111418193757000_8.24.3008.17",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.87.8.3082"
    },
    {
      "key": "2021111418193757000_8.24.3008.18",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(84)90372-6"
    },
    {
      "key": "2021111418193757000_8.24.3008.19",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.1853200"
    },
    {
      "key": "2021111418193757000_8.24.3008.20",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/S0960-9822(00)00061-0"
    },
    {
      "key": "2021111418193757000_8.24.3008.21",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.89.12.5482"
    },
    {
      "key": "2021111418193757000_8.24.3008.22",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(86)90066-8"
    },
    {
      "key": "2021111418193757000_8.24.3008.23",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(87)90614-3"
    },
    {
      "key": "2021111418193757000_8.24.3008.24",
      "doi-asserted-by": "crossref",
      "first-page": "247",
      "DOI": "10.1016/0167-4781(93)90122-T",
      "article-title": "Non-snRNP protein splicing factors.",
      "volume": "1173",
      "year": "1993",
      "journal-title": "Biochim. Biophys. Acta"
    },
    {
      "key": "2021111418193757000_8.24.3008.25",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.76.11.5495"
    },
    {
      "key": "2021111418193757000_8.24.3008.26",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(92)90556-R"
    },
    {
      "key": "2021111418193757000_8.24.3008.27",
      "doi-asserted-by": "publisher",
      "DOI": "10.1101/gad.8.9.1071"
    },
    {
      "key": "2021111418193757000_8.24.3008.28",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(92)90557-S"
    },
    {
      "key": "2021111418193757000_8.24.3008.29",
      "doi-asserted-by": "publisher",
      "DOI": "10.1101/gad.5.12b.2534"
    },
    {
      "key": "2021111418193757000_8.24.3008.30",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.1589782"
    },
    {
      "key": "2021111418193757000_8.24.3008.31",
      "unstructured": "Moore, M.J., C.C. Query, and P.A Sharp. 1993. Splicing of precursors to mRNA by the spliceosome. In The RNA world (ed. R.F. Gesteland and J.F. Atkins), pp. 303\u00e2\u0080\u0093357. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York."
    },
    {
      "key": "2021111418193757000_8.24.3008.32",
      "doi-asserted-by": "publisher",
      "DOI": "10.1021/bi00169a019"
    },
    {
      "key": "2021111418193757000_8.24.3008.33",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(94)90563-0"
    },
    {
      "key": "2021111418193757000_8.24.3008.34",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(87)90564-2"
    },
    {
      "key": "2021111418193757000_8.24.3008.35",
      "doi-asserted-by": "publisher",
      "DOI": "10.1101/gad.5.7.1224"
    },
    {
      "key": "2021111418193757000_8.24.3008.36",
      "unstructured": "Query, C.C. 1990. \u00e2\u0080\u009cAnalysis of RNA-protein interactions by identification of an RNA recognition motif within the U1 snRNA 70K autoantigen and definition of a protein-binding domain of U1 RNA,\u00e2\u0080\u009d Ph.D. thesis, Duke University, Durham, NC."
    },
    {
      "key": "2021111418193757000_8.24.3008.37",
      "doi-asserted-by": "publisher",
      "DOI": "10.1101/gad.8.5.587"
    },
    {
      "key": "2021111418193757000_8.24.3008.38",
      "doi-asserted-by": "publisher",
      "DOI": "10.1101/gad.2.10.1268"
    },
    {
      "key": "2021111418193757000_8.24.3008.39",
      "doi-asserted-by": "publisher",
      "DOI": "10.1126/science.2973660"
    },
    {
      "key": "2021111418193757000_8.24.3008.40",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(88)90509-0"
    },
    {
      "key": "2021111418193757000_8.24.3008.41",
      "doi-asserted-by": "publisher",
      "DOI": "10.1016/0092-8674(89)90296-1"
    },
    {
      "key": "2021111418193757000_8.24.3008.42",
      "doi-asserted-by": "crossref",
      "first-page": "1209",
      "DOI": "10.1002/j.1460-2075.1991.tb08062.x",
      "article-title": "The yeast branchpoint sequence is not required for the formation of a stable U1 snRNP pre-mRNA complex and is recognized in the absence of U2 snRNA.",
      "volume": "10",
      "year": "1991",
      "journal-title": "EMBO J."
    },
    {
      "key": "2021111418193757000_8.24.3008.43",
      "first-page": "2994",
      "article-title": "Direct interactions between pre-mRNA and six U2 small nuclear ribonucleoproteins during spliceosome assembly.",
      "volume": "14",
      "year": "1994",
      "journal-title": "Mol. Cell. Biol."
    },
    {
      "key": "2021111418193757000_8.24.3008.44",
      "doi-asserted-by": "crossref",
      "unstructured": "Takahashi, K. and S. Moore. 1982. Ribonuclease T1. In The enzymes (ed. P.D. Boyer), pp. 435\u00e2\u0080\u0093468. Academic Press, New York.",
      "DOI": "10.1016/S1874-6047(08)60285-1"
    },
    {
      "key": "2021111418193757000_8.24.3008.45",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.91.2.594"
    },
    {
      "key": "2021111418193757000_8.24.3008.46",
      "doi-asserted-by": "crossref",
      "first-page": "5467",
      "DOI": "10.1016/S0021-9258(19)57239-8",
      "article-title": "Autoantibodies reactive with U1 RNA and initiator methionine transfer RNA.",
      "volume": "261",
      "year": "1986",
      "journal-title": "J. Biol. Chem."
    },
    {
      "key": "2021111418193757000_8.24.3008.47",
      "doi-asserted-by": "publisher",
      "DOI": "10.1101/gad.3.10.1553"
    },
    {
      "key": "2021111418193757000_8.24.3008.48",
      "doi-asserted-by": "publisher",
      "DOI": "10.1101/gad.6.12b.2542"
    },
    {
      "key": "2021111418193757000_8.24.3008.49",
      "doi-asserted-by": "crossref",
      "first-page": "207",
      "DOI": "10.1002/j.1460-2075.1991.tb07937.x",
      "article-title": "Biochemical characterization of U2 snRNP auxiliary factor: An essential pre-mRNA splicing factor with a novel intranuclear distribution.",
      "volume": "10",
      "year": "1991",
      "journal-title": "EMBO J."
    },
    {
      "key": "2021111418193757000_8.24.3008.50",
      "doi-asserted-by": "publisher",
      "DOI": "10.1038/355609a0"
    },
    {
      "key": "2021111418193757000_8.24.3008.51",
      "doi-asserted-by": "publisher",
      "DOI": "10.1101/gad.3.10.1545"
    },
    {
      "key": "2021111418193757000_8.24.3008.52",
      "doi-asserted-by": "publisher",
      "DOI": "10.1073/pnas.86.8.2752"
    }
  ],
  "container-title": "Genes &amp; Development",
  "original-title": [],
  "language": "en",
  "link": [
    {
      "URL": "https://syndication.highwire.org/content/doi/10.1101/gad.8.24.3008",
      "content-type": "unspecified",
      "content-version": "vor",
      "intended-application": "similarity-checking"
    }
  ],
  "deposited": {
    "date-parts": [
      [
        2021,
        11,
        15
      ]
    ],
    "date-time": "2021-11-15T02:28:37Z",
    "timestamp": 1636943317000
  },
  "score": 1,
  "resource": {
    "primary": {
      "URL": "http://genesdev.cshlp.org/lookup/doi/10.1101/gad.8.24.3008"
    }
  },
  "subtitle": [],
  "short-title": [],
  "issued": {
    "date-parts": [
      [
        1994,
        12,
        15
      ]
    ]
  },
  "references-count": 52,
  "journal-issue": {
    "issue": "24",
    "published-print": {
      "date-parts": [
        [
          1994,
          12,
          15
        ]
      ]
    }
  },
  "alternative-id": [
    "10.1101/gad.8.24.3008"
  ],
  "URL": "http://dx.doi.org/10.1101/gad.8.24.3008",
  "relation": {},
  "ISSN": [
    "0890-9369",
    "1549-5477"
  ],
  "subject": [],
  "container-title-short": "Genes Dev.",
  "published": {
    "date-parts": [
      [
        1994,
        12,
        15
      ]
    ]
  }
}