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Yeast Two‐Hybrid and One‐Hybrid Screenings Identify Regulators of hsp70 Gene Expression
Author(s) -
Saito Youhei,
Nakagawa Takanobu,
Kakihana Ayana,
Nakamura Yoshia,
Nabika Tomomi,
Kasai Michihiro,
Takamori Mai,
Yamagishi Nobuyuki,
Kuga Takahisa,
Hatayama Takumi,
Nakayama Yuji
Publication year - 2016
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.25517
Subject(s) - hsp70 , gene knockdown , activator (genetics) , luciferase , biology , hsf1 , heat shock protein , gene expression , microbiology and biotechnology , gene , reporter gene , biochemistry , transfection
The mammalian stress protein Hsp105β, which is specifically expressed during mild heat shock and localizes to the nucleus, induces the major stress protein Hsp70. In the present study, we performed yeast two‐hybrid and one‐hybrid screenings to identify the regulators of Hsp105β‐mediated hsp70 gene expression. Six and two proteins were detected as Hsp105β‐ and hsp70 promoter‐binding proteins, respectively. A luciferase reporter gene assay revealed that hsp70 promoter activation is enhanced by the transcriptional co‐activator AF9 and splicing mediator SNRPE, but suppressed by the coiled‐coil domain‐containing protein CCDC127. Of these proteins, the knockdown of SNRPE suppressed the expression of Hsp70 irrespective of the presence of Hsp105β, indicating that SNRPE essentially functions as a transcriptional activator of hsp70 gene expression. The overexpression of HSP70 in tumor cells has been associated with cell survival and drug resistance. We here identified novel regulators of Hsp70 expression in stress signaling and also provided important insights into Hsp70‐targeted anti‐cancer therapy. J. Cell. Biochem. 117: 2109–2117, 2016. © 2016 Wiley Periodicals, Inc.