The inhibition of CMV promoter by heat shock factor 4b is regulated by Daxx.
Heat shock factor 4 (Hsf4b) has been identified as a novel cataractogenic protein whose mutation has been closely associated with hereditary cataracts in humans and animals. It acts both as a transcription activator and a transcription inhibitor in the regulation of its downstream targets during lens development. However, the signaling factors that regulate Hsf4b transcription activity are still not completely defined. Here, we found that Hsf4b, not Hsf4a (another isoform of Hsf4), acts as the inhibitor of CMV promoter as well as the activator of Hsp25 in the Hsf4-/- mouse lens epithelial cell line (mLEC/hsf4-/-). Hsf4b inhibits CMV-promoter activity by directly binding to TTCC (HSE motif) at 173-176bps in the CMV promoter. The phosphorylation of Hsf4b/S299 in the PDSM motif, which is absent in Hsf4a, participates in the negative regulation of the CMV promoter. The transcriptional inhibition of Hsf4b is associated with transcriptional inhibitor Daxx. Hsf4b can interact and co-localize with Daxx in the nucleus, and their association is regulated by the phosphorylation of Hsf4b/S299. In addition, we found that Hsf4a and Hsf1 were also associated with Daxx. However, in contrast to activating Hsf1, Daxx can repress Hsf4b-induced expression of Hsp25 in the mLEC/hsf4-/- cell line. Our results demonstrate that the transcription-inhibitory function of Hsf4b is regulated by the phosphorylation of Hsf4b/S299 and phosphorylation-dependent association with Daxx.
Zhang
Jun
J
Key Laboratory of Molecular and Cellular Immunology, Henan University School of Medicine, Kaifeng, China.
Hu
Yan-Zhong
YZ
Xueli
Li
L
Li
Shulian
S
Wang
Mingli
M
Kong
Xiangyin
X
Li
Tao
T
Shen
Peifen
P
Ma
Yuanfang
Y
D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D000818:Animals; D002386:Cataract / Q000235:genetics*; D002461:Cell Line, Transformed; D056970:Co-Repressor Proteins; D003587:Cytomegalovirus / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D015536:Down-Regulation / Q000235:genetics; D004847:Epithelial Cells / Q000378:metabolism* / Q000473:pathology; D055785:Gene Knockdown Techniques; D057809:HEK293 Cells; D000076249:Heat Shock Transcription Factors; D006801:Humans; D007908:Lens, Crystalline / Q000473:pathology; D051379:Mice; D018832:Molecular Chaperones; D009154:Mutation / Q000235:genetics; D009687:Nuclear Proteins / Q000378:metabolism*; D010766:Phosphorylation; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
D048868:Adaptor Proteins, Signal Transducing; D056970:Co-Repressor Proteins; C106985:DAXX protein, human; D004268:DNA-Binding Proteins; C102948:HSF4 protein, human; D000076249:Heat Shock Transcription Factors; C484462:Hsf4 protein, mouse; D018832:Molecular Chaperones; D009687:Nuclear Proteins; D014157:Transcription Factors