hElp3 directly modulates the expression of HSP70 gene in HeLa cells via HAT activity.
Human Elongator complex, which plays a key role in transcript elongation in vitro assay, is incredibly similar in either components or function to its yeast counterpart. However, there are only a few studies focusing on its target gene characterization in vivo. We studied the effect of down-regulation of the human elongation protein 3 (hELP3) on the expression of HSP70 through antisense strategy. Transfecting antisense plasmid p1107 into HeLa cells highly suppressed hELP3 expression, and substantially reduced expression of HSP70 mRNA and protein. Furthermore, chromatin immunoprecipitation assay (ChIP Assay) revealed that hElp3 participates in the transcription elongation of HSPA1A in HeLa cells. Finally, complementation and ChIP Assay in yeast showed that hElp3 can not only complement the growth and slow activation of HSP70 (SSA3) gene transcription, but also directly regulates the transcription of SSA3. On the contrary, these functions are lost when the HAT domain is deleted from hElp3. These data suggest that hElp3 can regulate the transcription of HSP70 gene, and the HAT domain of hElp3 is essential for this function. These findings now provide novel insights and evidence of the functions of hELP3 in human cells.
College of Life Science, Henan Normal University, Xinxiang, China. lifen2001@yahoo.com
Fen
F
Li
Jixian
J
Ma
Yu
Y
Ma
Yanyan
Y
Hu
Shujuan
S
Tian
Richard E
RE
White
Guichun
G
Han
D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D047369:Chromatin Immunoprecipitation; D017931:DNA Primers; D015536:Down-Regulation; D005816:Genetic Complementation Test; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics*; D006367:HeLa Cells; D051548:Histone Acetyltransferases / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D009419:Nerve Tissue Proteins / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction
D017931:DNA Primers; D018840:HSP70 Heat-Shock Proteins; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; C538729:ELP3 protein, human; D051548:Histone Acetyltransferases