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Specific interaction of the TCERG1 FF4–6 tandem repeat domains with RNA polymerase II requires simultaneous phosphorylation at Ser2, Ser5 and Ser7 of the CTD
Author(s) -
LIU JIANGXIN,
Fan Shilong,
Lee ChulJin,
Greenleaf Arno L,
Zhou Pei
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.1015.2
Subject(s) - ctd , rna polymerase ii , heptad repeat , rna splicing , transcription (linguistics) , eukaryotic transcription , phosphorylation , chemistry , tandem repeat , microbiology and biotechnology , alternative splicing , computational biology , biology , rna , biochemistry , promoter , peptide sequence , gene , messenger rna , gene expression , geology , oceanography , linguistics , philosophy , genome
The human transcription elongation regulator TCERG1 physically couples transcription elongation and splicing events by interacting with splicing factors through its N‐terminal WW domains and the hyperphosphorylated C‐terminal domain (CTD) of RNA polymerase II (RNAPII) through its C‐terminal FF domains. Here, we report biochemical and structural characterization of the C‐terminal three FF domains (FF4–6) of TCERG1, revealing a rigid integral domain structure of the tandem FF domains that interacts with the hyperphosphorylated CTD (PCTD). Using peptide column binding assays and NMR titrations, we show that binding of FF4–6 toward the PCTD requires simultaneous phosphorylation at Ser2, Ser5 and Ser7 positions within the Y1S2P3T4S5P6S7 heptad repeats. Such a sequence‐specific PCTD recognition is achieved through CTD‐docking sites on FF4 and FF5 of TCERG1, but not FF6. In vivo yeast cell extract pull down assay demonstrates the essential role of Ser7P. Our study presents the first example of a nuclear factor requiring all three phosphoSer marks within the heptad repeat of the CTD for high‐affinity binding and provides a molecular interpretation for the biochemical connection between the Ser7P enrichment and co‐transcriptional splicing events. This research was supported by NIH grants GM079376 (to P.Z.) and GM040505 (A.L.G.).