title: Specific interaction of the transcription elongation regulator TCERG1 with RNA polymerase II requires simultaneous phosphorylation at Ser2, Ser5, and Ser7 within the carboxyl-terminal domain repeat.
abstract: 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 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 repeat that interacts with the hyperphosphorylated CTD (PCTD). Although FF4 and FF5 adopt a classical FF domain fold containing three orthogonally packed α helices and a 310 helix, FF6 contains an additional insertion helix between α1 and α2. The formation of the integral tandem FF4-6 repeat is achieved by merging the last helix of the preceding FF domain and the first helix of the following FF domain and by direct interactions between neighboring FF domains. Using peptide column binding assays and NMR titrations, we show that binding of the FF4-6 tandem repeat to the PCTD requires simultaneous phosphorylation at Ser(2), Ser(5), and Ser(7) positions within two consecutive Y(1)S(2)P(3)T(4)S(5)P(6)S(7) heptad repeats. Such a sequence-specific PCTD recognition is achieved through CTD-docking sites on FF4 and FF5 of TCERG1 but not FF6. Our study presents the first example of a nuclear factor requiring all three phospho-Ser marks within the heptad repeat of the CTD for high affinity binding and provides a molecular interpretation for the biochemical connection between the Ser(7) phosphorylation enrichment in the CTD of the transcribing RNA polymerase II over introns and co-transcriptional splicing events.
authors: Department of Biochemistry, Duke University Medical Center, Durham, North Carolina 27710, USA.
authors: Jiangxin
authors: J
authors: Liu
authors: Shilong
authors: S
authors: Fan
authors: Chul-Jin
authors: CJ
authors: Lee
authors: Arno L
authors: AL
authors: Greenleaf
authors: Pei
authors: P
authors: Zhou
mesh_terms: D006801:Humans; D019906:Nuclear Magnetic Resonance, Biomolecular; D010766:Phosphorylation / Q000502:physiology; D011485:Protein Binding / Q000502:physiology; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012319:RNA Polymerase II / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D020449:Repetitive Sequences, Amino Acid; D012694:Serine / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D035602:Transcriptional Elongation Factors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: C108440:TCERG1 protein, human; D035602:Transcriptional Elongation Factors; D012694:Serine; D012319:RNA Polymerase II