The interaction of RNA polymerase II with the adenovirus-2 major late promoter is precluded by phosphorylation of the C-terminal domain of subunit IIa - PubMed (original) (raw)
. 1992 May 25;267(15):10500-6.
Affiliations
- PMID: 1316903
Free article
The interaction of RNA polymerase II with the adenovirus-2 major late promoter is precluded by phosphorylation of the C-terminal domain of subunit IIa
J D Chesnut et al. J Biol Chem. 1992.
Free article
Abstract
Mammalian RNA polymerase II contains at the C terminus of its largest subunit an unusual domain consisting of 52 tandem repeats of the consensus sequence Tyr-Ser-Pro-Thr-Ser-Pro-Ser. The phosphorylation of this domain is thought to play an important role in the transition of RNA polymerase II from a preinitiation complex to an elongating complex. The unphosphorylated form of RNA polymerase II is designated IIA, whereas the phosphorylated form is designated IIO. In an effort to determine the consequence of C-terminal domain phosphorylation on complex formation, 32P-labeled RNA polymerases IIA and IIO were prepared and examined for their ability to form a stable preinitiation complex on the adenovirus-2 major late promoter in the presence of a reconstituted HeLa cell transcription extract. Preinitiation complexes were formed in the absence of ATP and purified from free RNA polymerase II by chromatography on Sepharose CL-4B. The state of phosphorylation of the largest subunit was monitored by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the transcriptional activity was determined by assaying specific transcript formation upon the addition of nucleotides and a competing DNA template. RNA polymerase IIA was recovered in transcriptionally active complexes in reactions in which the input enzyme was RNA polymerase IIA. In reactions with RNA polymerase IIO as the input enzyme, no IIO was recovered in excluded fractions that normally contain preinitiation complex. In reactions with equimolar amounts of RNA polymerases IIO and IIA, purified preinitiation complexes contained almost exclusively RNA polymerase HA. These results support the idea that RNA polymerase II containing an unphosphorylated C-terminal domain preferentially associates with the adenovirus-2 major late promoter. The state of phosphorylation of the C-terminal domain can, therefore, directly influence preinitiation complex formation. We also report here the presence of an activity in HeLa cell extracts that catalyzes dephosphorylation of the C-terminal domain, thereby converting RNA polymerase IIO to IIA. This C-terminal domain phosphatase is specific in that it does not catalyze the dephosphorylation of a serine residue phosphorylated by casein kinase II. The presence of a C-terminal domain phosphatase in in vitro transcription reactions containing RNA polymerase IIO results in the formation of RNA polymerase IIA. This RNA polymerase IIA associates preferentially with preinitiation complexes.
Similar articles
- Phosphorylation of the C-terminal domain of RNA polymerase II.
Dahmus ME. Dahmus ME. Biochim Biophys Acta. 1995 Apr 4;1261(2):171-82. doi: 10.1016/0167-4781(94)00233-s. Biochim Biophys Acta. 1995. PMID: 7711060 Review. - Phosphorylation and functions of the RNA polymerase II CTD.
Phatnani HP, Greenleaf AL. Phatnani HP, et al. Genes Dev. 2006 Nov 1;20(21):2922-36. doi: 10.1101/gad.1477006. Genes Dev. 2006. PMID: 17079683 Review.
Cited by
- RNA polymerase II is aberrantly phosphorylated and localized to viral replication compartments following herpes simplex virus infection.
Rice SA, Long MC, Lam V, Spencer CA. Rice SA, et al. J Virol. 1994 Feb;68(2):988-1001. doi: 10.1128/JVI.68.2.988-1001.1994. J Virol. 1994. PMID: 8289400 Free PMC article. - Serine/threonine phosphorylation regulates binding of C hnRNP proteins to pre-mRNA.
Mayrand SH, Dwen P, Pederson T. Mayrand SH, et al. Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7764-8. doi: 10.1073/pnas.90.16.7764. Proc Natl Acad Sci U S A. 1993. PMID: 8356082 Free PMC article. - Analysis of the role of TFIIE in basal transcription and TFIIH-mediated carboxy-terminal domain phosphorylation through structure-function studies of TFIIE-alpha.
Ohkuma Y, Hashimoto S, Wang CK, Horikoshi M, Roeder RG. Ohkuma Y, et al. Mol Cell Biol. 1995 Sep;15(9):4856-66. doi: 10.1128/MCB.15.9.4856. Mol Cell Biol. 1995. PMID: 7651404 Free PMC article. - ICP22 and the UL13 protein kinase are both required for herpes simplex virus-induced modification of the large subunit of RNA polymerase II.
Long MC, Leong V, Schaffer PA, Spencer CA, Rice SA. Long MC, et al. J Virol. 1999 Jul;73(7):5593-604. doi: 10.1128/JVI.73.7.5593-5604.1999. J Virol. 1999. PMID: 10364308 Free PMC article. - A motif shared by TFIIF and TFIIB mediates their interaction with the RNA polymerase II carboxy-terminal domain phosphatase Fcp1p in Saccharomyces cerevisiae.
Kobor MS, Simon LD, Omichinski J, Zhong G, Archambault J, Greenblatt J. Kobor MS, et al. Mol Cell Biol. 2000 Oct;20(20):7438-49. doi: 10.1128/MCB.20.20.7438-7449.2000. Mol Cell Biol. 2000. PMID: 11003641 Free PMC article.