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PMID: 7957047 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Enhanced phosphorylation of the C-terminal domain of RNA polymerase II upon serum stimulation of quiescent cells: possible involvement of MAP kinases.

The EMBO journal ·Vol. 13 ·No. 20 ·1994-10-17 ·Pages 4787-97

Dubois MF, Nguyen VT, Dahmus ME, Pagès G, Pouysségur J, Bensaude O

Abstract

The largest subunit of RNA polymerase (RNAP) II contains at it C-terminus an unusual domain comprising tandem repeats of the consensus sequence Tyr-Ser-Pro-Thr-Ser-Pro-Ser. This C-terminal domain (CTD) can undergo phosphorylation at multiple sites giving rise to a form of the enzyme designated RNAP IIO. The unphosphorylated form is designated RNAP IIA. The largest subunits of RNAPs IIO and IIA are designated IIo and IIa, respectively. In quiescent NIH 3T3 fibroblasts, subunits IIo and IIa are present in comparable amounts. Upon serum stimulation, the amount of subunit IIo increases markedly and remains elevated for several hours. The increase of subunit IIo also occurs in transcription-inhibited cells and, therefore, is not a consequence of serum-activated transcription. This observation suggests that serum stimulation activates a CTD kinase and/or inhibits a CTD phosphatase. This hypothesis is supported by the finding that serum stimulates phosphorylation of a beta-galactosidase-CTD fusion protein expressed in these cells. Furthermore, an enhanced CTD kinase activity was discovered in lysates from serum-stimulated fibroblasts and was found to copurify with MAP kinases on a Mono Q column and to bind to anti-MAP kinase antibodies. The idea that MAP kinases phosphorylate the CTD in vivo is supported by the observation that subunit IIa, but not subunit IIb which lacks the CTD, is phosphorylated at multiple sites by purified MAP kinase. Consequently, the MAP kinases are a new class of CTD kinases which appear to be involved in the phosphorylation of RNAP II following serum stimulation. This phosphorylation may contribute to the transcriptional activation of serum-stimulated genes.

MeSH Terms
3T3 Cells Amino Acid Sequence Animals Blood Cross Reactions Enzyme Activation Mice Mitogens Molecular Sequence Data Phosphorylation Precipitin Tests Protein Kinases/metabolism RNA Polymerase II/metabolism Recombinant Fusion Proteins/metabolism Transcription, Genetic beta-Galactosidase/metabolism
Chemicals
Mitogens Recombinant Fusion Proteins Protein Kinases carboxy-terminal domain kinase RNA Polymerase II beta-Galactosidase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Dubois M F
Laboratoire de Génétique Moléculaire, URA CNRS no. 1302, Paris, France.
Nguyen V T
Dahmus M E
Pagès G
Pouysségur J
Bensaude O
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1994-10-17
Pages
4787-97
Language
English
Region
England
NLM ID
8208664
PMCID
PMC395417
Subset
IM
Grants
PHS HHS · 33300 · United States
External Links
PubMed source PMC Full Text View journal details
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