title: The integrator complex recognizes a new double mark on the RNA polymerase II carboxyl-terminal domain.
abstract: The carboxyl-terminal domain (CTD) of the largest subunit of RNA polymerase II (pol II) comprises multiple tandem repeats of the heptapeptide Tyr(1)-Ser(2)-Pro(3)-Thr(4)-Ser(5)-Pro(6)-Ser(7). This unusual structure serves as a platform for the binding of factors required for expression of pol II-transcribed genes, including the small nuclear RNA (snRNA) gene-specific Integrator complex. The pol II CTD specifically mediates recruitment of Integrator to the promoter of snRNA genes to activate transcription and direct 3' end processing of the transcripts. Phosphorylation of the CTD and a serine in position 7 are necessary for Integrator recruitment. Here, we have further investigated the requirement of the serines in the CTD heptapeptide and their phosphorylation for Integrator binding. We show that both Ser(2) and Ser(7) of the CTD are required and that phosphorylation of these residues is necessary and sufficient for efficient binding. Using synthetic phosphopeptides, we have determined the pattern of the minimal Ser(2)/Ser(7) double phosphorylation mark required for Integrator to interact with the CTD. This novel double phosphorylation mark is a new addition to the functional repertoire of the CTD code and may be a specific signal for snRNA gene expression.
authors.affiliation: Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, United Kingdom.
authors.forename: Sylvain
authors.initials: S
authors.lastname: Egloff
authors.forename: Sylwia Anna
authors.initials: SA
authors.lastname: Szczepaniak
authors.forename: Martin
authors.initials: M
authors.lastname: Dienstbier
authors.forename: Alice
authors.initials: A
authors.lastname: Taylor
authors.forename: Sophie
authors.initials: S
authors.lastname: Knight
authors.forename: Shona
authors.initials: S
authors.lastname: Murphy
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D015153:Blotting, Western; D002467:Cell Nucleus / Q000201:enzymology; D005982:Glutathione Transferase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D009842:Oligopeptides / Q000737:chemistry / Q000235:genetics; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000302:isolation & purification / Q000378:metabolism; D012319:RNA Polymerase II / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D012342:RNA, Small Nuclear / Q000235:genetics; D012694:Serine / Q000302:isolation & purification / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D009842:Oligopeptides; D010446:Peptide Fragments; D012342:RNA, Small Nuclear; D012694:Serine; D005982:Glutathione Transferase; D017346:Protein Serine-Threonine Kinases; D012319:RNA Polymerase II
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