Structure of an mRNA Capping Enzyme Bound to the Phosphorylated Carboxy-Terminal Domain of RNA Polymerase II

ABSTRACT The cotranscriptional placement of the 7-methylguanosine cap on pre-mRNA is mediated by recruitment of capping enzyme to the phosphorylated carboxy-terminal domain (CTD) of RNA polymerase II. Immunoblotting suggests that the capping enzyme guanylyltransferase (Ceg1) is stabilized in vivo by its interaction with the CTD and that serine 5, the major site of phosphorylation within the CTD heptamer consensus YSPTSPS, is particularly important. We sought to identify the CTD kinase responsible for capping enzyme targeting. The candidate kinases Kin28-Ccl1, CTDK1, and Srb10-Srb11 can each phosphorylate a glutathione S-transferase–CTD fusion protein such that capping enzyme can bind in vitro. However, kin28 mutant alleles cause reduced Ceg1 levels in vivo and exhibit genetic interactions with a mutant ceg1 allele, whilesrb10 or ctk1 deletions do not. Therefore, only the TFIIH-associated CTD kinase Kin28 appears necessary for proper capping enzyme targeting in vivo. Interestingly, levels of the polyadenylation factor Pta1 are also reduced in kin28 mutants, while several other polyadenylation factors remain stable. Pta1 in yeast extracts binds specifically to the phosphorylated CTD, suggesting that this interaction may mediate coupling of polyadenylation and transcription.

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Faculty Opinions recommendation of Recognition of RNA polymerase II carboxy-terminal domain by 3'-RNA-processing factors.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1020073.229427 ◽

2004 ◽

Author(s):

Benoit Coulombe

Keyword(s):

Rna Polymerase ◽

Rna Polymerase Ii ◽

Rna Processing ◽

Terminal Domain ◽

Carboxy Terminal Domain ◽

Carboxy Terminal ◽

Processing Factors

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What’s all the phos about? Insights into the phosphorylation state of the RNA polymerase II C-terminal domain via mass spectrometry

RSC Chemical Biology ◽

10.1039/d1cb00083g ◽

2021 ◽

Author(s):

Blase Matthew LeBlanc ◽

Rosamaria Yvette Moreno ◽

Edwin Escobar ◽

Mukesh Kumar Venkat Ramani ◽

Jennifer S Brodbelt ◽

...

Keyword(s):

Rna Polymerase ◽

Rna Polymerase Ii ◽

Phosphorylation State ◽

Terminal Domain ◽

Carboxy Terminal Domain ◽

Protein Encoding ◽

Small Nuclear Rnas ◽

Rnap Ii ◽

Carboxy Terminal ◽

Encoding Genes

RNA polymerase II (RNAP II) is one of the primary enzymes responsible for expressing protein-encoding genes and some small nuclear RNAs. The enigmatic carboxy-terminal domain (CTD) of RNAP II and...

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Study on the association between CTD peptides and zinc(ii)-dipicolylamine appended beta-cyclodextrin

RSC Advances ◽

10.1039/c5ra12374g ◽

2015 ◽

Vol 5 (98) ◽

pp. 80434-80440 ◽

Cited By ~ 1

Author(s):

Saihui Zhang ◽

Yantao Shi ◽

Wei Wang ◽

Zhi Yuan

Keyword(s):

Rna Polymerase ◽

Rna Polymerase Ii ◽

Terminal Domain ◽

Carboxy Terminal Domain ◽

Beta Cyclodextrin ◽

Carboxy Terminal

Association between zinc(ii)-dipicolylamine appended beta-cyclodextrin and CTD (carboxy-terminal domain of RNA polymerase II) peptides with different phosphorylation patterns was studied by ITC and NMR.

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Synthetically engineeredrpb1alleles altering RNA polymerase II carboxy terminal domain phosphorylation induce discrete morphogenetic defects inSchizosaccharomyces pombe

Communicative & Integrative Biology ◽

10.4161/cib.23954 ◽

2013 ◽

Vol 6 (3) ◽

pp. e23954 ◽

Cited By ~ 2

Author(s):

Kyle Hoffman ◽

Haeyoung Yoo ◽

Jim Karagiannis

Keyword(s):

Rna Polymerase ◽

Rna Polymerase Ii ◽

Terminal Domain ◽

Carboxy Terminal Domain ◽

Carboxy Terminal

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Heat shock-induced alterations in phosphorylation of the largest subunit of RNA polymerase II as revealed by monoclonal antibodies CC-3 and MPM-2

Biochemistry and Cell Biology ◽

10.1139/o99-037 ◽

1999 ◽

Vol 77 (4) ◽

pp. 367-374 ◽

Cited By ~ 3

Author(s):

Sébastien B Lavoie ◽

Alexandra L Albert ◽

Alain Thibodeau ◽

Michel Vincent

Keyword(s):

Heat Shock ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Transcriptional Activity ◽

Phosphorylation Sites ◽

Stress Genes ◽

Terminal Domain ◽

Carboxy Terminal Domain ◽

Heat Shocks ◽

Carboxy Terminal

The phosphorylation of the carboxy-terminal domain of the largest subunit of RNA polymerase II plays an important role in the regulation of transcriptional activity and is also implicated in pre-mRNA processing. Different stresses, such as a heat shock, induce a marked alteration in the phosphorylation of this domain. The expression of stress genes by RNA polymerase II, to the detriment of other genes, could be attributable to such modifications of the phosphorylation sites. Using two phosphodependent antibodies recognizing distinct hyperphosphorylated forms of RNA polymerase II largest subunit, we studied the phosphorylation state of the subunit in different species after heat shocks of varying intensities. One of these antibodies, CC-3, preferentially recognizes the carboxy-terminal domain of the largest subunit under normal conditions, but its reactivity is diminished during stress. In contrast, the other antibody used, MPM-2, demonstrated a strong reactivity after a heat shock in most species studied. Therefore, CC-3 and MPM-2 antibodies discriminate between phosphoisomers that may be functionally different. Our results further indicate that the pattern of phosphorylation of RNA polymerase II in most species varies in response to environmental stress.Key words: RNA polymerase II, heat shock, phosphorylation, CC-3, MPM-2.

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