Identification of a novel gene product, Sertoli cell gene with a zinc finger domain, that is important for FSH activation of testicular Sertoli cells
Identification of a novel gene product, Sertoli cell gene with a zinc finger domain, that is important for FSH activation of testicular Sertoli cells
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DOI：
10.1210/en.143.2.426
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发表时间：
2002-02-01
期刊：
影响因子：
4.8
通讯作者：
Skinner, MK
中科院分区：
文献类型：
--
作者：
Chaudhary, J;Skinner, MK
支持细胞为精子发生过程提供细胞结构支持和微环境。从支持细胞中分离到一个新的、广泛表达的cDNA克隆，命名为含锌指结构域的支持细胞基因（SERZ）。一个显着的同源性SERZ被发现与小鼠基因组序列，建议存在至少10个外显子。在cDNA的5 '端的开放阅读框，称为SERZ-α，具有隐蔽的碱性螺旋-环-螺旋（bHLH）结构域，但没有起始密码子。当将起始密码子工程化到cDNA的5 '端时，获得SERZ-α的体外翻译产物。最长的第二开放阅读框，在距离5 '端304 bp处具有ATG起始位点，编码308个氨基酸的SERZ-β多肽。SERZ-β的基序分析和BLAST搜索显示与保守锌指蛋白的DHHC结构域具有显著的同源性。在SERZ-β多肽序列中观察到许多潜在的磷酸化位点。SERZ-β的长的5 '-非翻译区促使了对两种潜在的替代多肽产物SERZ-α和SERZ-β的研究。在总支持细胞蛋白的Western印迹分析中，用SERZ-β和5 '端开放阅读框SERZ-α的特异性抗体检测SERZ-α和SERZ-β蛋白。通过cDNA的体外翻译也证实了SERZ-β多肽的存在，但在不存在工程化起始密码子的情况下，SERZ-α不在体外翻译。SERZ mRNA的表达模式在所有检查的组织中观察到。通过北方印迹分析测定的SERZ的转录物大小为约2.7 kb。SERZ的反义寡核苷酸被发现不影响基础水平的转铁蛋白启动子激活，但显着阻断FSH诱导的转铁蛋白启动子激活。SERZ mRNA表达不受FSH处理支持细胞培养物的调节。总之，一个新的基因产物，SERZ，被确定，似乎有一个作用，在维持支持细胞分化的功能和介导FSH的行动。SERZ的翻译可能产生两种基因产物;然而，含有锌指结构域的SERZ-β可能是SERZ基因的主要产物。
Sertoli cells provide the cytoarchitectural support and microenvironment necessary for the process of spermatogenesis. A novel, ubiquitously expressed cDNA clone was isolated from Sertoli cells and termed Sertoli cell gene with a zinc finger domain (SERZ). A significant homology of SERZ was found with a mouse genomic sequence that suggested the presence of at least 10 exons. An open reading frame at the 5'-end of the cDNA, termed SERZ-alpha, had a cryptic basic helix-loop-helix (bHLH) domain, but no start codon. When a start codon was engineered into the 5'-end of the cDNA, an in vitro translation product of SERZ-alpha was obtained. The longest second open reading frame with an ATG start site at 304 bp from the 5'-end coded for a 308-amino acid SERZ-beta polypeptide. Motif analysis and BLAST search of SERZ-beta showed significant homology to the DHHC domain of conserved zinc finger proteins. A number of potential phosphorylation sites were observed in the SERZ-beta polypeptide sequence. The long 5'-untranslated region of SERZ-beta prompted an investigation of both potential alternate polypeptide products, SERZ-alpha and SERZ-beta. Both SERZ-alpha and SERZ-beta proteins were detected with specific antibodies to SERZ-beta and the 5'-end open reading frame SERZ-alpha in a Western blot analysis of total Sertoli cell proteins. The presence of the SERZ-beta polypeptide was also confirmed by in vitro translation of the cDNA, but SERZ-alpha was not translated in vitro in the absence of an engineered start codon. The expression pattern of SERZ mRNA was observed in all tissues examined. The transcript size of SERZ as determined by Northern blot analysis is approximately 2.7 kb. An antisense oligonucleotide to SERZ was found not to influence basal levels of transferrin promoter activation, but significantly blocked FSH-induced transferrin promoter activation. SERZ mRNA expression was not regulated by FSH treatment of Sertoli cell cultures. In summary, a novel gene product, SERZ, was identified that appears to have a role in maintaining Sertoli cell differentiated functions and mediating FSH actions. Translation of SERZ may give rise to two gene products; however, the SERZ-beta containing the zinc finger domain is probably the principal product of the SERZ gene.