DOI: 10.1101/gad.2038011. The RNA polymerase II C-terminal domain promotes splicing activation through recruitment of a U2AF65

The RNA polymerase II C-terminal domain promotes splicing activation through recruitment of a U2AF65–Prp19 complex

10.1101/gad.2038011

Crossref journal-article

Cold Spring Harbor Laboratory

Genes & Development (246)

Abstract

Pre-mRNA splicing is frequently coupled to transcription by RNA polymerase II (RNAPII). This coupling requires the C-terminal domain of the RNAPII largest subunit (CTD), although the underlying mechanism is poorly understood. Using a biochemical complementation assay, we previously identified an activity that stimulates CTD-dependent splicing in vitro. We purified this activity and found that it consists of a complex of two well-known splicing factors: U2AF65 and the Prp19 complex (PRP19C). We provide evidence that both U2AF65 and PRP19C are required for CTD-dependent splicing activation, that U2AF65 and PRP19C interact both in vitro and in vivo, and that this interaction is required for activation of splicing. Providing the link to the CTD, we show that U2AF65 binds directly to the phosphorylated CTD, and that this interaction results in increased recruitment of U2AF65 and PRP19C to the pre-mRNA. Our results not only provide a mechanism by which the CTD enhances splicing, but also describe unexpected interactions important for splicing and its coupling to transcription.

Bibliography

David, C. J., Boyne, A. R., Millhouse, S. R., & Manley, J. L. (2011). The RNA polymerase II C-terminal domain promotes splicing activation through recruitment of a U2AF65âPrp19 complex. Genes & Development, 25(9), 972â983.

Authors 4

Charles J. David (first)
Alex R. Boyne (additional)
Scott R. Millhouse (additional)
James L. Manley (additional)

References 67 Referenced 163

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{'key': '2021111618511363000_25.9.972.55', 'first-page': '1883', 'article-title': 'The yeast PRP19 protein is not tightly associated with small nuclear RNAs, but appears to associate with the spliceosome after binding of U2 to the pre-mRNA and prior to formation of the functional spliceosome', 'volume': '13', 'year': '1993', 'journal-title': 'Mol Cell Biol'} / Mol Cell Biol / The yeast PRP19 protein is not tightly associated with small nuclear RNAs, but appears to associate with the spliceosome after binding of U2 to the pre-mRNA and prior to formation of the functional spliceosome (1993)
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10.1128/MCB.20.21.8290-8301.2000
10.1101/gad.10.11.1356

Dates

Type	When
Created	14 years, 3 months ago (May 2, 2011, 10:41 a.m.)
Deposited	3 years, 9 months ago (Nov. 16, 2021, 9:52 p.m.)
Indexed	17 hours, 26 minutes ago (Aug. 21, 2025, 1:06 p.m.)
Issued	14 years, 3 months ago (May 1, 2011)
Published	14 years, 3 months ago (May 1, 2011)
Published Online	14 years, 3 months ago (May 2, 2011)
Published Print	14 years, 3 months ago (May 1, 2011)

Funders 0

None

BibTeX

@article{David_2011, title={The RNA polymerase II C-terminal domain promotes splicing activation through recruitment of a U2AF65–Prp19 complex}, volume={25}, ISSN={1549-5477}, url={http://dx.doi.org/10.1101/gad.2038011}, DOI={10.1101/gad.2038011}, number={9}, journal={Genes & Development}, publisher={Cold Spring Harbor Laboratory}, author={David, Charles J. and Boyne, Alex R. and Millhouse, Scott R. and Manley, James L.}, year={2011}, month=may, pages={972–983} }

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    "10.1101/gad.2038011"
  ],
  "URL": "http://dx.doi.org/10.1101/gad.2038011",
  "relation": {},
  "ISSN": [
    "0890-9369",
    "1549-5477"
  ],
  "subject": [],
  "container-title-short": "Genes Dev.",
  "published": {
    "date-parts": [
      [
        2011,
        5,
        1
      ]
    ]
  }
}