Oncogenic function of SCCRO5/DCUN1D5 requires its Neddylation E3 activity and nuclear localization.
Oncogenic function of SCCRO5/DCUN1D5 requires its Neddylation E3 activity and nuclear localization.
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SCCRO5/DCUN1D5的致癌功能需要其Neddylation E3活性和核定位。
DOI：
10.1158/1078-0432.ccr-13-1252
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发表时间：
2014-01-15
期刊：
影响因子：
--
通讯作者：
Singh B
中科院分区：
文献类型：
--
作者：
Bommeljé CC;Weeda VB;Huang G;Shah K;Bains S;Buss E;Shaha M;Gönen M;Ghossein R;Ramanathan SY;Singh B
确定SCCRO 5（又名DCUN 1D 5）促进肿瘤发生的机制。通过实时PCR和Western印迹分析，在203个随机选择的原发性癌组织样本、匹配的组织学正常组织和细胞系中评估SCCRO 5 mRNA和蛋白表达。SCCRO 5过表达与生存相关。在选定的癌细胞系中评估SCCRO 5敲低对生存力的影响。进行结构-功能研究以确定与neddylation组分结合、neddylation促进活性和转化所需的SCCRO 5残基。在口腔和肺鳞状细胞癌中，SCCRO 5 mRNA水平与蛋白水平相对应，过表达与疾病特异性生存率降低相关。通过RNAi敲低SCCRO 5导致具有高内源性水平的癌细胞活力的选择性降低，表明存在癌基因成瘾。SCCRO 5促进cullin neddylation，同时保持参与泛素和泛素样蛋白缀合的保守反应持续进行性范例，将其确立为neddylation E3的组分。体外Neddylation活性的增强需要Neddylation（PONY）结构域的参与，而不需要核定位序列（NLS）结构域的参与。相反，NIH-3 T3细胞的转化需要NLS结构域和PONY结构域。我们的数据表明，SCCRO 5具有致癌潜力，它赋予其致癌性作为neddylation E3的一个组成部分。SCCRO 5的Neddylation活性和核定位对其体内功能非常重要。
To determine mechanisms by which SCCRO5 (aka DCUN1D5) promotes oncogenesis. SCCRO5 mRNA and protein expression were assessed in 203 randomly selected primary cancer tissue samples, matched histologically normal tissues, and cell lines by use of real-time PCR and Western blot analysis. SCCRO5 overexpression was correlated with survival. The effect of SCCRO5 knockdown on viability was assessed in selected cancer cell lines. Structure-function studies were performed to determine the SCCRO5 residues required for binding to the neddylation components, for neddylation-promoting activity, and for transformation. In oral and lung squamous cell carcinomas, SCCRO5 mRNA levels corresponded with protein levels and overexpression correlated with decreased disease-specific survival. Knockdown of SCCRO5 by RNAi resulted in a selective decrease in the viability of cancer cells with high endogenous levels, suggesting the presence of oncogene addiction. SCCRO5 promoted cullin neddylation while maintaining conserved reaction processivity paradigms involved in ubiquitin and ubiquitin-like protein conjugation, establishing it as a component of the neddylation E3. Neddylation activities in vitro required the potentiating of neddylation (PONY) domain but not the nuclear localization sequence (NLS) domain. In contrast, both the NLS domain and the PONY domain were required for transformation of NIH-3T3 cells. Our data suggest that SCCRO5 has oncogenic potential and that it imparts its oncogenicity as a component of the neddylation E3. Neddylation activity and nuclear localization of SCCRO5 are important for its in vivo function.