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Sanada, Sayoko; Maekawa, Masashi; Tate, Sota; Nakaoka, Hiroki; Fujisawa, Yasuhiro; Sayama, Koji; Higashiyama, Shigeki
Biochemical and biophysical research communications, 04/2023, Volume: 651Journal Article
Speckle-type pox virus and zinc finger (POZ) protein (SPOP), a substrate recognition receptor for the cullin-3/RING ubiquitin E3 complex, leads to the ubiquitination of >40 of its target substrates. Since a variety of point mutations in the substrate-binding domain of SPOP have been identified in cancers, including prostate and endometrial cancers, the pathological roles of those cancer-associated SPOP mutants have been extensively elucidated. In this study, we evaluated the cellular functions of wild-type SPOP in non-cancerous human keratinocyte-derived HaCaT cells expressing wild-type SPOP gene. SPOP knockdown using siRNA in HaCaT cells dramatically reduced cell growth and arrested their cell cycles at G1/S phase. The expression of DNA replication licensing factors CDT1 and CDC6 in HaCaT cells drastically decreased on SPOP knockdown as their translation was inhibited. CDT1 and CDC6 downregulation induced p21 expression without p53 activation. Our results suggest that SPOP is essential for DNA replication licensing in non-cancerous keratinocyte HaCaT cells. •SPOP (speckle-type POZ protein) is essential for DNA synthesis and cell proliferation in HaCaT cells.•Depletion of SPOP reduces protein expression of DNA replication licensing factors, CDT1 and CDC6, in HaCaT cells.•Translation of CDT1 and CDC6 is remarkably inhibited by SPOP knockdown in HaCaT cells.
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