© The Rockefeller University Press,
0021-9525/1997//5 $5.00
The Journal of Cell Biology, Volume 136, Number 1,
, 1997 5-18
A Functional Interaction between the Carboxy-Terminal Domain of RNA Polymerase II and Pre-mRNA Splicing
Lei Du* and
Stephen L. Warren
Department of Pathology and * Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06510
In the preceding study we found that Sm snRNPs and SerArg (SR) family proteins co-immunoprecipitate with Pol II molecules containing a hyperphosphorylated CTD (Kim et al., 1997). The association between Pol IIo and splicing factors is maintained in the absence of pre-mRNA, and the polymerase need not be transcriptionally engaged (Kim et al., 1997). The latter findings led us to hypothesize that a phosphorylated form of the CTD interacts with pre-mRNA splicing components in vivo. To test this idea, a nested set of CTD-derived proteins was assayed for the ability to alter the nuclear distribution of splicing factors, and to interfere with splicing in vivo. Proteins containing heptapeptides 1-52 (CTD52), 1-32 (CTD32), 1-26 (CTD26), 1-13 (CTD13), 1-6 (CTD6), 1-3 (CTD3), or 1 (CTD1) were expressed in mammalian cells. The CTD-derived proteins become phosphorylated in vivo, and accumulate in the nucleus even though they lack a conventional nuclear localization signal. CTD52 induces a selective reorganization of splicing factors from discrete nuclear domains to the diffuse nucleoplasm, and significantly, it blocks the accumulation of spliced, but not unspliced, human β-globin transcripts. The extent of splicing factor disruption, and the degree of inhibition of splicing, are proportional to the number of heptapeptides added to the protein. The above results indicate a functional interaction between Pol II's CTD and pre-mRNA splicing.
Abbreviations used in this paper: CBs, coiled bodies; CTD, carboxy-terminal domain of RNA polymerase II; MIG, mitotic interchromatin granule cluster; nt, nucleotide; Pol IIa, unphosphorylated largest RNA polymerase II subunit; Pol II LS, largest subunit of RNA polymerase II; Pol IIo, hyperphosphorylated largest RNA polymerase II subunit; RT-PCR, reverse transcription-polymerase chain reaction; Sm sn RNP, Smith antigen-containing small nuclear ribonucleoprotein; SR, serine-arginine dipeptide repeat motif.
The work was supported by the Council for Tobacco Research (#3881) and National Institutes of Health (K08-CA01339) to S.L. Warren.
Please address all correspondence to S.L. Warren, Brady Memorial Laboratories, Room B117, Department of Pathology, Yale University School of Medicine, P.O. Box 208023, New Haven, CT 06520-8023. Tel.: (203) 737-2247. Fax: (203) 785-7303. E-mail: stephen.warren{at}yale.edu

CiteULike
Complore
Connotea
Del.icio.us
Digg
Facebook
Reddit
Technorati
Twitter What's this?
This article has been cited by other articles:
-
McAlinden, A., Liang, L., Mukudai, Y., Imamura, T., Sandell, L. J.
(2007). Nuclear Protein TIA-1 Regulates COL2A1 Alternative Splicing and Interacts with Precursor mRNA and Genomic DNA. J. Biol. Chem.
282: 24444-24454
[Abstract]
[Full Text]
-
Lacadie, S. A., Tardiff, D. F., Kadener, S., Rosbash, M.
(2006). In vivo commitment to yeast cotranscriptional splicing is sensitive to transcription elongation mutants. Genes Dev.
20: 2055-2066
[Abstract]
[Full Text]
-
Sanchez-Alvarez, M., Goldstrohm, A. C., Garcia-Blanco, M. A., Sune, C.
(2006). Human Transcription Elongation Factor CA150 Localizes to Splicing Factor-Rich Nuclear Speckles and Assembles Transcription and Splicing Components into Complexes through Its Amino and Carboxyl Regions.. Mol. Cell. Biol.
26: 4998-5014
[Abstract]
[Full Text]
-
KORNBLIHTT, A. R., DE LA MATA, M., FEDEDA, J. P., MUNOZ, M. J., NOGUES, G.
(2004). Multiple links between transcription and splicing. RNA
10: 1489-1498
[Abstract]
[Full Text]
-
Metz, A., Soret, J., Vourc'h, C., Tazi, J., Jolly, C.
(2004). A key role for stress-induced satellite III transcripts in the relocalization of splicing factors into nuclear stress granules. J. Cell Sci.
117: 4551-4558
[Abstract]
[Full Text]
-
Fomenkov, A., Huang, Y.-P., Topaloglu, O., Brechman, A., Osada, M., Fomenkova, T., Yuriditsky, E., Trink, B., Sidransky, D., Ratovitski, E.
(2003). p63{alpha} Mutations Lead to Aberrant Splicing of Keratinocyte Growth Factor Receptor in the Hay-Wells Syndrome. J. Biol. Chem.
278: 23906-23914
[Abstract]
[Full Text]
-
Pardee, T. S., Ghazy, M. A., Ponticelli, A. S.
(2003). Yeast and Human RNA Polymerase II Elongation Complexes: Evidence for Functional Differences and Postinitiation Recruitment of Factors. Eukaryot Cell
2: 318-327
[Abstract]
[Full Text]
-
Matsuda, K.-i., Ochiai, I., Nishi, M., Kawata, M.
(2002). Colocalization and Ligand-Dependent Discrete Distribution of the Estrogen Receptor (ER){alpha} and ER{beta}. Mol. Endocrinol.
16: 2215-2230
[Abstract]
[Full Text]
-
Robert, F., Blanchette, M., Maes, O., Chabot, B., Coulombe, B.
(2002). A Human RNA Polymerase II-containing Complex Associated with Factors Necessary for Spliceosome Assembly. J. Biol. Chem.
277: 9302-9306
[Abstract]
[Full Text]
-
Goldstrohm, A. C., Albrecht, T. R., Sune, C., Bedford, M. T., Garcia-Blanco, M. A.
(2001). The Transcription Elongation Factor CA150 Interacts with RNA Polymerase II and the Pre-mRNA Splicing Factor SF1. Mol. Cell. Biol.
21: 7617-7628
[Abstract]
[Full Text]
-
Fong, N., Bentley, D. L.
(2001). Capping, splicing, and 3' processing are independently stimulated by RNA polymerase II: different functions for different segments of the CTD. Genes Dev.
15: 1783-1795
[Abstract]
[Full Text]
-
Adams, D. J., van der Weyden, L., Mayeda, A., Stamm, S., Morris, B. J., Rasko, J. E.J.
(2001). ZNF265--a novel spliceosomal protein able to induce alternative splicing. JCB
154: 25-32
[Abstract]
[Full Text]
-
Zeng, C., Berget, S. M.
(2000). Participation of the C-Terminal Domain of RNA Polymerase II in Exon Definition during Pre-mRNA Splicing. Mol. Cell. Biol.
20: 8290-8301
[Abstract]
[Full Text]
-
Schroeder, S. C., Schwer, B., Shuman, S., Bentley, D.
(2000). Dynamic association of capping enzymes with transcribing RNA polymerase II. Genes Dev.
14: 2435-2440
[Abstract]
[Full Text]
-
Garber, M. E., Mayall, T. P., Suess, E. M., Meisenhelder, J., Thompson, N. E., Jones, K. A.
(2000). CDK9 Autophosphorylation Regulates High-Affinity Binding of the Human Immunodeficiency Virus Type 1 Tat-P-TEFb Complex to TAR RNA. Mol. Cell. Biol.
20: 6958-6969
[Abstract]
[Full Text]
-
Hirose, Y., Manley, J. L.
(2000). RNA polymerase II and the integration of nuclear events. Genes Dev.
14: 1415-1429
[Full Text]
-
Lewis, J. D., Tollervey, D.
(2000). Like Attracts Like: Getting RNA Processing Together in the Nucleus. Science
288: 1385-1389
[Abstract]
[Full Text]
-
Yang, L., Embree, L. J., Hickstein, D. D.
(2000). TLS-ERG Leukemia Fusion Protein Inhibits RNA Splicing Mediated by Serine-Arginine Proteins. Mol. Cell. Biol.
20: 3345-3354
[Abstract]
[Full Text]
-
Misteli, T
(2000). Cell biology of transcription and pre-mRNA splicing: nuclear architecture meets nuclear function. J. Cell Sci.
113: 1841-1849
[Abstract]
-
Morris, D. P., Phatnani, H. P., Greenleaf, A. L.
(1999). Phospho-Carboxyl-Terminal Domain Binding and the Role of a Prolyl Isomerase in Pre-mRNA 3'-End Formation. J. Biol. Chem.
274: 31583-31587
[Abstract]
[Full Text]
-
Douziech, M., Forget, D., Greenblatt, J., Coulombe, B.
(1999). Topological Localization of the Carboxyl-Terminal Domain of RNA Polymerase II in the Initiation Complex. J. Biol. Chem.
274: 19868-19873
[Abstract]
[Full Text]
-
Jolly, C., Vourc'h, C., Robert-Nicoud, M., Morimoto, R. I.
(1999). Intron-independent Association of Splicing Factors with Active Genes. JCB
145: 1133-1143
[Abstract]
[Full Text]
-
Hirose, Y., Tacke, R., Manley, J. L.
(1999). Phosphorylated RNA polymerase II stimulates pre-mRNA splicing. Genes Dev.
13: 1234-1239
[Abstract]
[Full Text]
-
Lai, M.-C., Teh, B. H., Tarn, W.-Y.
(1999). A Human Papillomavirus E2 Transcriptional Activator. THE INTERACTIONS WITH CELLULAR SPLICING FACTORS AND POTENTIAL FUNCTION IN PRE-mRNA PROCESSING. J. Biol. Chem.
274: 11832-11841
[Abstract]
[Full Text]
-
Snaar, S. P., Vincent, M., Dirks, R. W.
(1999). RNA Polymerase II Localizes at Sites of Human Cytomegalovirus Immediate-early RNA Synthesis and Processing. J. Histochem. Cytochem.
47: 245-254
[Abstract]
[Full Text]
-
Collas, P, Liang, M., Vincent, M, Alestrom, P
(1999). Active transgenes in zebrafish are enriched in acetylated histone H4 and dynamically associate with RNA Pol II and splicing complexes. J. Cell Sci.
112: 1045-1054
[Abstract]
-
Abdellatif, M., Packer, S. E., Michael, L. H., Zhang, D., Charng, M. J., Schneider, M. D.
(1998). A Ras-Dependent Pathway Regulates RNA Polymerase II Phosphorylation in Cardiac Myocytes: Implications for Cardiac Hypertrophy. Mol. Cell. Biol.
18: 6729-6736
[Abstract]
[Full Text]
-
Huang, S., Deerinck, T. J., Ellisman, M. H., Spector, D. L.
(1998). The Perinucleolar Compartment and Transcription. JCB
143: 35-47
[Abstract]
[Full Text]
-
Wolff, T., O'Neill, R. E., Palese, P.
(1998). NS1-Binding Protein (NS1-BP): a Novel Human Protein That Interacts with the Influenza A Virus Nonstructural NS1 Protein Is Relocalized in the Nuclei of Infected Cells. J. Virol.
72: 7170-7180
[Abstract]
[Full Text]
-
Baurén, G., Belikov, S., Wieslander, L.
(1998). Transcriptional termination in the Balbiani ring 1 gene is closely coupled to 3'-end formation and excision of the 3'-terminal intron. Genes Dev.
12: 2759-2769
[Abstract]
[Full Text]
-
Hampsey, M.
(1998). Molecular Genetics of the RNA Polymerase II General Transcriptional Machinery. Microbiol. Mol. Biol. Rev.
62: 465-503
[Abstract]
[Full Text]
-
Politz, J. C., Browne, E. S., Wolf, D. E., Pederson, T.
(1998). Intranuclear diffusion and hybridization state of oligonucleotides measured by fluorescence correlation spectroscopy in living cells. Proc. Natl. Acad. Sci. USA
95: 6043-6048
[Abstract]
[Full Text]
-
Patturajan, M., Wei, X., Berezney, R., Corden, J. L.
(1998). A Nuclear Matrix Protein Interacts with the Phosphorylated C-Terminal Domain of RNA Polymerase II. Mol. Cell. Biol.
18: 2406-2415
[Abstract]
[Full Text]
-
Patturajan, M., Schulte, R. J., Sefton, B. M., Berezney, R., Vincent, M., Bensaude, O., Warren, S. L., Corden, J. L.
(1998). Growth-related Changes in Phosphorylation of Yeast RNA Polymerase II. J. Biol. Chem.
273: 4689-4694
[Abstract]
[Full Text]
-
Loyer, P, Trembley, J., Lahti, J., Kidd, V.
(1998). The RNP protein, RNPS1, associates with specific isoforms of the p34cdc2-related PITSLRE protein kinase in vivo. J. Cell Sci.
111: 1495-1506
[Abstract]
-
Powers, C. A., Mathur, M., Raaka, B. M., Ron, D., Samuels, H. H.
(1998). TLS (Translocated-in-Liposarcoma) Is a High-Affinity Interactor for Steroid, Thyroid Hormone, and Retinoid Receptors. Mol. Endocrinol.
12: 4-18
[Abstract]
[Full Text]
-
MCCRACKEN, S., ROSONINA, E., FONG, N., SIKES, M., BEYER, A., O'HARE, K., SHUMAN, S., BENTLEY, D.
(1998). Role of RNA Polymerase II Carboxy-terminal Domain in Coordinating Transcription with RNA Processing. Cold Spring Harb Symp Quant Biol
63: 301-310
[Abstract]
-
Neugebauer, K. M., Roth, M. B.
(1997). Transcription units as RNA processing units. Genes Dev.
11: 3279-3285
[Full Text]
-
Cho, E.-J., Takagi, T., Moore, C. R., Buratowski, S.
(1997). mRNA capping enzyme is recruited to the transcription complex by phosphorylation of the RNA polymerase II carboxy-terminal domain. Genes Dev.
11: 3319-3326
[Abstract]
[Full Text]
-
Grondin, B., Cote, F., Bazinet, M., Vincent, M., Aubry, M.
(1997). Direct Interaction of the KRAB/Cys2-His2 Zinc Finger Protein ZNF74 with a Hyperphosphorylated Form of the RNA Polymerase II Largest Subunit. J. Biol. Chem.
272: 27877-27885
[Abstract]
[Full Text]
-
Caceres, J. F., Misteli, T., Screaton, G. R., Spector, D. L., Krainer, A. R.
(1997). Role of the Modular Domains of SR Proteins in Subnuclear Localization and Alternative Splicing Specificity. JCB
138: 225-238
[Abstract]
[Full Text]
-
Lee, J. M., Greenleaf, A. L.
(1997). Modulation of RNA Polymerase II Elongation Efficiency by C-terminal Heptapeptide Repeat Domain Kinase I. J. Biol. Chem.
272: 10990-10993
[Abstract]
[Full Text]
-
Morris, D. P., Greenleaf, A. L.
(2000). The Splicing Factor, Prp40, Binds the Phosphorylated Carboxyl-terminal Domain of RNA Polymerase II. J. Biol. Chem.
275: 39935-39943
[Abstract]
[Full Text]
-
Stoss, O., Olbrich, M., Hartmann, A. M., Konig, H., Memmott, J., Andreadis, A., Stamm, S.
(2001). The STAR/GSG Family Protein rSLM-2 Regulates the Selection of Alternative Splice Sites. J. Biol. Chem.
276: 8665-8673
[Abstract]
[Full Text]