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Transcriptional properties of mammalian Elongin A and its role in stress response (944.1)
Author(s) -
Inoue Makoto,
Kawauchi Junya,
Fukuda Mizue,
Uchida Yohei,
Yasukawa Takashi,
Conaway Ronald,
Conaway Joan,
Aso Teijiro,
Kitajima Shigetaka
Publication year - 2014
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.28.1_supplement.944.1
Subject(s) - rna polymerase ii , transcription (linguistics) , transcription factor , elongation factor , biology , microbiology and biotechnology , mutant , gene expression , immunoprecipitation , gene , rna , genetics , promoter , ribosome , linguistics , philosophy
Transcriptional elongation factor Elongin A exhibits an activity to repress the temporary pause of RNA polymerase II (RNAPII) on the DNA template. The role of Elongin A in RNAPII transcription in mammalian cells has not, however, been clearly established. In this study, we investigate the function of Elongin A in RNAPII transcription. First, we examined immunoprecipitation and immunostaining. These showed that Elongin A binds to the phosphorylated form of RNAPII (IIO) at both Ser5 and Ser2 and partially colocalized with the RNAPIIO and newly transcribed RNA. Significantly, Western blotting and qRT‐PCR showed that Elongin A was required for the maximal induction of stress response gene ATF3 and p21 . ChIP assay showed that Elongin A occupancy was increased on ATF3 , p21, c‐Myc and c‐jun genes after doxorubicin treatment. Furthermore, we performed re‐introduction assay of various mutants of Elongin A. It indicated that expressions of ATF3 and p21 gene were dependent on elongation activity of Elongin A. By contrast, polyubiquitination of RNAPII was dependent on ubiqutination activity of Elongin A. Taken together, our data provide the evidence that mammalian Elongin A is involved in the elongation phase of transcription in vivo and plays a crucial role in response to stress stimuli. Furthermore, our data indicated that two activities of Elongin A are independent of one another in response to stress.