SPOP is essential for DNA replication licensing through maintaining translation of CDT1 and CDC6 in HaCaT cells
SPOP is essential for DNA replication licensing through maintaining translation of CDT1 and CDC6 in HaCaT cells
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DOI：
10.1016/j.bbrc.2023.02.012
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发表时间：
2023-02-13
影响因子：
3.1
通讯作者：
Higashiyama，Shigeki
中科院分区：
文献类型：
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作者：
Sanada，Sayoko;Maekawa，Masashi;Higashiyama，Shigeki
斑点型痘病毒和锌指（POZ）蛋白（SPOP）是cullin-3/RING泛素E3复合物的底物识别受体，导致其靶底物中>40种的泛素化。由于SPOP的底物结合结构域中的多种点突变已在癌症（包括前列腺癌和子宫内膜癌）中被鉴定，这些癌症相关的SPOP突变体的病理作用已被广泛阐明。在本研究中，我们在表达野生型SPOP基因的非癌人角质形成细胞来源的HaCaT细胞中评估了野生型SPOP的细胞功能，使用siRNA在HaCaT细胞中敲低SPOP显著地降低了细胞生长并将其细胞周期阻滞在G1/S期。DNA复制许可因子CDT 1和CDC 6在HaCaT细胞中的表达在SPOP敲低时显著降低，因为它们的翻译被抑制。CDT 1和CDC 6下调诱导p21表达而无p53活化。我们的研究结果表明，SPOP是必不可少的非癌性角质形成细胞HaCaT细胞的DNA复制许可。
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-typeSPOPgene.SPOPknockdown 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 onSPOPknockdown 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.