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Studies of Schizosaccharomyces pombe TFIIE indicate conformational and functional changes in RNA polymerase II at transcription initiation
Author(s) -
Hayashi Kazuhiro,
Watanabe Tomomichi,
Tanaka Aki,
Furumoto Tadashi,
SatoTsuchiya Chiaki,
Kimura Makoto,
Yokoi Masayuki,
Ishihama Akira,
Hanaoka Fumio,
Ohkuma Yoshiaki
Publication year - 2005
Publication title -
genes to cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.912
H-Index - 115
eISSN - 1365-2443
pISSN - 1356-9597
DOI - 10.1111/j.1365-2443.2005.00833.x
Subject(s) - transcription factor ii e , biology , transcription factor ii a , rna polymerase ii , transcription factor ii f , general transcription factor , transcription preinitiation complex , transcription factor ii d , microbiology and biotechnology , heterotetramer , schizosaccharomyces pombe , transcription (linguistics) , transcription factor ii h , transcription factor ii b , protein subunit , transcription factor , biochemistry , promoter , rna , transcriptional regulation , rna polymerase , gene , mutant , gene expression , linguistics , philosophy
The general transcription factor TFIIE plays essential roles in transcription by RNA polymerase II (PolII). Despite recent progress, the elucidation of its precise mechanisms including biological functions awaits further characterization. We report the isolation and characterization of Schizosaccharomyces pombe TFIIE (spTFIIE). Like human and other eukaryotic TFIIE proteins, spTFIIE consists of α and β subunits and the genes encoding both subunits are essential for viability. Chromatin immunoprecipitation assays demonstrated that spTFIIE localizes to promoters in vivo . Mutational analysis of the C‐terminal basic helix‐loop region of TFIIEβ, which is involved in the transition from transcription initiation to elongation, revealed that transcription‐defective mutants affected in this region are also cold sensitive. The spTFIIEβ subunit binds both spTFIIEβ and spTFIIEα but spTFIIEα binds only spTFIIEβ. These results indicate that TFIIE forms an α 2 β 2 heterotetramer in which two αβ heterodimers are connected via β subunits. Further analysis of binding specificities showed that spTFIIEβ binds the Rpb2 and Rpb12 subunits of PolII, whereas spTFIIEα predominantly binds Rpb5, which is located at the clamp region and changes conformation upon transcription initiation.