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PMID: 24478330 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

3' end formation of pre-mRNA and phosphorylation of Ser2 on the RNA polymerase II CTD are reciprocally coupled in human cells.

Genes & development ·Vol. 28 ·No. 4 ·2014-02-15 ·Pages 342-56

Davidson L, Muniz L, West S

Abstract

3' end formation of pre-mRNAs is coupled to their transcription via the C-terminal domain (CTD) of RNA polymerase II (Pol II). Nearly all protein-coding transcripts are matured by cleavage and polyadenylation (CPA), which is frequently misregulated in disease. Understanding how transcription is coordinated with CPA in human cells is therefore very important. We found that the CTD is heavily phosphorylated on Ser2 (Ser2p) at poly(A) (pA) signals coincident with recruitment of the CstF77 CPA factor. Depletion of the Ser2 kinase Cdk12 impairs Ser2p, CstF77 recruitment, and CPA, strongly suggesting that the processes are linked, as they are in budding yeast. Importantly, we additionally show that the high Ser2p signals at the 3' end depend on pA signal function. Down-regulation of CPA results in the loss of a 3' Ser2p peak, whereas a new peak is formed when CPA is induced de novo. Finally, high Ser2p signals are generated by Pol II pausing, which is a well-known feature of pA site recognition. Thus, a reciprocal relationship between early steps in pA site processing and Ser2p ensures efficient 3' end formation.

Keywords
C-terminal domain Cdk12 RNA polymerase II Ser2 cleavage and polyadenylation pausing
MeSH Terms
Cyclin-Dependent Kinases/genetics,metabolism Gene Expression Regulation Humans Phosphorylation Protein Structure, Tertiary RNA Polymerase II/metabolism RNA Precursors/chemistry,genetics,metabolism Serine/metabolism
Chemicals
RNA Precursors Serine CDK12 protein, human Cyclin-Dependent Kinases RNA Polymerase II
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Davidson Lee
Wellcome Trust Centre for Cell Biology, Edinburgh EH9 3JR, United Kingdom.
Muniz Lisa
West Steven
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Article Info
Journal
Genes & development
Abbr.
Genes Dev
ISSN
1549-5477
DOI
10.1101/gad.231274.113
Published
2014-02-15
Epub
2014-00-29
Pages
342-56
Language
English
Region
United States
NLM ID
8711660
PMCID
PMC3937513
Subset
IM
Grants
Wellcome Trust · 088499/Z/09/2 · United Kingdom
Wellcome Trust · 092076 · United Kingdom
External Links
PubMed source DOI: 10.1101/gad.231274.113 PMC Full Text View journal details
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