This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kolodziej, P A Articles by Young, R A Search for Related Content PubMed PubMed Citation Articles by Kolodziej, P A Articles by Young, R A

 Previous Article  |  Next Article 

Mol Cell Biol. 1990 May; 10(5): 1915-1920

RNA polymerase II subunit composition, stoichiometry, and phosphorylation.

Whitehead Institute for Biomedical Research, Nine Cambridge Center, Massachusetts 02142.

ABSTRACT

RNA polymerase II subunit composition, stoichiometry, and phosphorylation were investigated in Saccharomyces cerevisiae by attaching an epitope coding sequence to a well-characterized RNA polymerase II subunit gene (RPB3) and by immunoprecipitating the product of this gene with its associated polypeptides. The immunopurified enzyme catalyzed alpha-amanitin-sensitive RNA synthesis in vitro. The 10 polypeptides that immunoprecipitated were identical in size and number to those previously described for RNA polymerase II purified by conventional column chromatography. The relative stoichiometry of the subunits was deduced from knowledge of the sequence of the subunits and from the extent of labeling with [35S]methionine. Immunoprecipitation from 32P-labeled cell extracts revealed that three of the subunits, RPB1, RPB2, and RPB6, are phosphorylated in vivo. Phosphorylated and unphosphorylated forms of RPB1 could be distinguished; approximately half of the RNA polymerase II molecules contained a phosphorylated RPB1 subunit. These results more precisely define the subunit composition and phosphorylation of a eucaryotic RNA polymerase II enzyme.

This article has been cited by other articles:

• Runner, V. M., Podolny, V., Buratowski, S. (2008). The Rpb4 Subunit of RNA Polymerase II Contributes to Cotranscriptional Recruitment of 3' Processing Factors. Mol. Cell. Biol. 28: 1883-1891 [Abstract] [Full Text]  
• Sampath, V., Balakrishnan, B., Verma-Gaur, J., Onesti, S., Sadhale, P. P. (2008). Unstructured N Terminus of the RNA Polymerase II Subunit Rpb4 Contributes to the Interaction of Rpb4{middle dot}Rpb7 Subcomplex with the Core RNA Polymerase II of Saccharomyces cerevisiae. J. Biol. Chem. 283: 3923-3931 [Abstract] [Full Text]  
• Gerber, J., Reiter, A., Steinbauer, R., Jakob, S., Kuhn, C.-D., Cramer, P., Griesenbeck, J., Milkereit, P., Tschochner, H. (2008). Site specific phosphorylation of yeast RNA polymerase I. Nucleic Acids Res 36: 793-802 [Abstract] [Full Text]  
• Martens, J. A., Wu, P.-Y. J., Winston, F. (2005). Regulation of an intergenic transcript controls adjacent gene transcription in Saccharomyces cerevisiae. Genes Dev. 19: 2695-2704 [Abstract] [Full Text]  
• Kaplan, C. D., Holland, M. J., Winston, F. (2005). Interaction between Transcription Elongation Factors and mRNA 3'-End Formation at the Saccharomyces cerevisiae GAL10-GAL7 Locus. J. Biol. Chem. 280: 913-922 [Abstract] [Full Text]  
• Sampath, V., Rekha, N., Srinivasan, N., Sadhale, P. (2003). The Conserved and Non-conserved Regions of Rpb4 Are Involved in Multiple Phenotypes in Saccharomyces cerevisiae. J. Biol. Chem. 278: 51566-51576 [Abstract] [Full Text]  
• Tan, Q., Prysak, M. H., Woychik, N. A. (2003). Loss of the Rpb4/Rpb7 Subcomplex in a Mutant Form of the Rpb6 Subunit Shared by RNA Polymerases I, II, and III. Mol. Cell. Biol. 23: 3329-3338 [Abstract] [Full Text]  
• Muratoglu, S., Georgieva, S., Papai, G., Scheer, E., Enunlu, I., Komonyi, O., Cserpan, I., Lebedeva, L., Nabirochkina, E., Udvardy, A., Tora, L., Boros, I. (2003). Two Different Drosophila ADA2 Homologues Are Present in Distinct GCN5 Histone Acetyltransferase-Containing Complexes. Mol. Cell. Biol. 23: 306-321 [Abstract] [Full Text]  
• Kimura, M., Suzuki, H., Ishihama, A. (2002). Formation of a Carboxy-Terminal Domain Phosphatase (Fcp1)/TFIIF/RNA Polymerase II (pol II) Complex in Schizosaccharomyces pombe Involves Direct Interaction between Fcp1 and the Rpb4 Subunit of pol II. Mol. Cell. Biol. 22: 1577-1588 [Abstract] [Full Text]  
• Honey, S., Schneider, B. L., Schieltz, D. M., Yates, J. R., Futcher, B. (2001). A novel multiple affinity purification tag and its use in identification of proteins associated with a cyclin-CDK complex. Nucleic Acids Res 29: e24-e24 [Abstract] [Full Text]  
• Tan, Q., Li, X., Sadhale, P. P., Miyao, T., Woychik, N. A. (2000). Multiple Mechanisms of Suppression Circumvent Transcription Defects in an RNA Polymerase Mutant. Mol. Cell. Biol. 20: 8124-8133 [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]  
• Todone, F., Weinzierl, R. O. J., Brick, P., Onesti, S. (2000). Crystal structure of RPB5, a universal eukaryotic RNA polymerase subunit and transcription factor interaction target. Proc. Natl. Acad. Sci. USA 97: 6306-6310 [Abstract] [Full Text]  
• Donaldson, I. M., Friesen, J. D. (2000). Zinc Stoichiometry of Yeast RNA Polymerase II and Characterization of Mutations in the Zinc-binding Domain of the Largest Subunit. J. Biol. Chem. 275: 13780-13788 [Abstract] [Full Text]  
• Sakurai, H., Mitsuzawa, H., Kimura, M., Ishihama, A. (1999). The Rpb4 Subunit of Fission Yeast Schizosaccharomyces pombe RNA Polymerase II Is Essential for Cell Viability and Similar in Structure to the Corresponding Subunits of Higher Eukaryotes. Mol. Cell. Biol. 19: 7511-7518 [Abstract] [Full Text]  
• Sheffer, A., Varon, M., Choder, M. (1999). Rpb7 Can Interact with RNA Polymerase II and Support Transcription during Some Stresses Independently of Rpb4. Mol. Cell. Biol. 19: 2672-2680 [Abstract] [Full Text]  
• Kershnar, E., Wu, S.-Y., Chiang, C. M. (1998). Immunoaffinity Purification and Functional Characterization of Human Transcription Factor IIH and RNA Polymerase II from Clonal Cell Lines That Conditionally Express Epitope-tagged Subunits of the Multiprotein Complexes. J. Biol. Chem. 273: 34444-34453 [Abstract] [Full Text]  
• Rosenheck, S., Choder, M. (1998). Rpb4, a Subunit of RNA Polymerase II, Enables the Enzyme To Transcribe at Temperature Extremes In Vitro. J. Bacteriol. 180: 6187-6192 [Abstract] [Full Text]  
• McNeil, J. B., Agah, H., Bentley, D. (1998). Activated transcription independent of the RNA polymerase II holoenzyme in budding yeast. Genes Dev. 12: 2510-2521 [Abstract] [Full Text]  
• Svetlov, V., Nolan, K., Burgess, R. R. (1998). Rpb3, Stoichiometry and Sequence Determinants of the Assembly into Yeast RNA Polymerase II in Vivo. J. Biol. Chem. 273: 10827-10830 [Abstract] [Full Text]  
• Kimura, M., Ishiguro, A., Ishihama, A. (1997). RNA Polymerase II Subunits 2, 3, and 11 Form a Core Subassembly with DNA Binding Activity. J. Biol. Chem. 272: 25851-25855 [Abstract] [Full Text]  
• Acker, J., de Graaff, M., Cheynel, I., Khazak, V., Kedinger, C., Vigneron, M. (1997). Interactions between the Human RNA Polymerase II Subunits. J. Biol. Chem. 272: 16815-16821 [Abstract] [Full Text]  
• Larkin, R. M., Guilfoyle, T. J. (1997). Reconstitution of Yeast and Arabidopsis RNA Polymerase alpha -like Subunit Heterodimers. J. Biol. Chem. 272: 12824-12830 [Abstract] [Full Text]  
• Ulmasov, T., Larkin, R. M., Guilfoyle, T. J. (1996). Association between 36- and 13.6-kDa alpha-Like Subunits of Arabidopsis thaliana RNA Polymerase II. J. Biol. Chem. 271: 5085-5094 [Abstract] [Full Text]  
• Weeks, J R, Hardin, S E, Shen, J, Lee, J M, Greenleaf, A L (1993). Locus-specific variation in phosphorylation state of RNA polymerase II in vivo: correlations with gene activity and transcript processing.. Genes Dev. 7: 2329-2344 [Abstract]  
• Bennett, M, Reed, R (1993). Correspondence between a mammalian spliceosome component and an essential yeast splicing factor. Science 262: 105-108 [Abstract]  
• Liao, S M, Taylor, I C, Kingston, R E, Young, R A (1991). RNA polymerase II carboxy-terminal domain contributes to the response to multiple acidic activators in vitro.. Genes Dev. 5: 2431-2440 [Abstract]