Cullin3 is a KLHL10-interacting protein preferentially expressed during late spermiogenesis
Cullin3 is a KLHL10-interacting protein preferentially expressed during late spermiogenesis
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DOI：
10.1095/biolreprod.105.045484
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发表时间：
2006-01-01
影响因子：
3.6
通讯作者：
Yan, W
中科院分区：
文献类型：
--
作者：
Wang, SH;Zheng, HL;Yan, W
Kelch-like 10 (KLHL10) 是 BTB（Bric-a-brac、Tramtrack 和 Broad-Complex）-kelch 蛋白超家族的成员，对于精子发生和男性生育能力至关重要。在使用酵母双杂交测定寻找 KLHL10 相互作用蛋白时，我们确定 Cullin3 (CUL3) 是多个 KLHL10 相互作用蛋白之一。酵母共转化测定表明，CUL3 结合 KLHL10 的 BTB/POZ 结构域。 Northern blot 和定量 RT-PCR 分析表明 Cul3 mRNA 优先在睾丸中表达。原位杂交分析将 Cul3 mRNA 定位于成年睾丸中的精子细胞。通过免疫荧光显微镜在伸长和伸长的精子细胞中检测到 CUL3 蛋白（步骤 10-16）。 CUL3 的表达模式类似于 KLHL10。 CUL3 与 KLHL10 共免疫沉淀，并且使用睾丸裂解物在 CUL3 免疫沉淀剂中也检测到 KLHL10。这些发现表明，KLHL10 与其他 BTB/kelch 蛋白一样，与 CUL3 相互作用，形成基于 CUL3 的泛素 E3 连接酶，该酶在睾丸中特异性发挥作用，在精子发生过程中介导蛋白质泛素化。
Kelch-like 10 (KLHL10) is a member of the BTB (Bric-a-brac, Tramtrack, and Broad-Complex)-kelch protein superfamily essential for spermiogenesis and male fertility. In a search for KLHL10-interacting proteins using a yeast two-hybrid assay, we identified Cullin3 (CUL3) as one of multiple KLHL10-interacting partners. Yeast cotransformation assays revealed that CUL3 bound the BTB/POZ domain of KLHL10. Northern blot and quantitative RT-PCR analyses demonstrated that Cul3 mRNA was preferentially expressed in the testis. in situ hybridization analysis localized Cul3 mRNA to spermatids in the adult testis. CUL3 protein was detected in elongating and elongated spermatids (steps 10-16) by immunofluorescent microscopy. The expression pattern of CUL3 resembles KLHL10. CUL3 was coimmunoprecipated with KLHL10, and KLHL10 was also detected in the CUL3 immunoprecipitants using testis lysates. These findings suggest that KLHL10, like other BTB/kelch proteins, interacts with CUL3 to form a CUL3-based ubiquitin E3 ligase that functions specifically in the testis to mediate protein ubiquitination during spermiogenesis.