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胱蛋白酶抑制剂相关的附睾精子发生基因的调节和功能 被引量:2

Cres (Cystatin-Related Epididymal Spermatogenic) Gene Regulation and Function
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摘要 CRES(cystatin relatedepididymalspermatogenic,胱蛋白酶抑制剂相关的附睾精子发生 )蛋白是胱蛋白酶抑制剂(cystatin)超家族中家族 2的一个亚类。然而 ,与cystatinC的广泛性表达不同 ,Cres基因只在分裂后的生殖细胞、附睾头部近端和腺垂体促性腺激素细胞中表达。cystatin对C1半胱氨酸蛋白酶抑制作用的发挥必须有 3个共有位点的参与 ,而CRES蛋白缺少其中的 2个位点。因此 ,CERS在生殖系统和神经内分泌系统中的功能也许是独特的和组织特异性的。本文概述了以下方面的研究 :①Cres基因启动子及其转录调节蛋白相关的可能对Cres的组织特异性表达有重要作用的DNA结合位点。②CRES蛋白的生物学功能。Norethern印迹法、凝胶转移分析和瞬时转染实验均表明 ,附睾和促性腺激素细胞中主要表达C EBP家族中的C EBPβ(CCAAT 增强子结合蛋白 ) ,且C EBPβ对于Cres基因在这两个组织中的高表达是必不可少的。另外 ,构建了表达氯霉素乙酰基转移酶 (CAT)报告基因的转基因小鼠 ,CAT报告基因由Cres基因 5′端 1 6kb的启动子所调控。分析显示 ,CATmRNA仅在生殖细胞中表达 ,说明Cres 5′端 1 6kb的侧翼区含有调控CAT在睾丸中表达的DNA序列 ,而缺乏指导CAT在附睾中表达的序列 ,或者此1 6kb的DNA片段存在Cres的负调节成分。最后 , The CRES (cystatin related epididymal spermatogenic) protein defines a new subgroup in the family 2 cystatins of the cystatin superfamily of cysteine protease inhibitors. However, unlike the ubiquitous expression of cystatin C, the Cres gene is preferentialy expressed in postmeiotic germ cells, the proximal caput epididymidis, and anterior pituitary gonadotrophs. Furthermore, CRES protein lacks two of the three consensus sites necessary for the cystatin inhibition of C1 cysteine proteases. Therefore, CRES may perform unique and tissue specific functions in the reproductive and neuroendocrine systems. In the present review, we describe our studies on: ①the Cres gene promoter and the transcriptional regulatory proteins and their associated DNA binding sites that may be important for tissue specific expression; and ②the biochemical function of CRES protein. In brief, Northern blot, gel shift analyses, and transient transfection assays demonstrated that the C/EBPβ (CCAAT/enhancer binding protein) transcription factor is the predominant C/EBP family member expressed in the epididymis and gonadotroph cells and is necessary for high levels of Cres expression in these two tissues. In other studies, analyses of transgenic mice expressing a CAT reporter gene driven by 1.6 kb of Cres promoter revealed CAT mRNA and protein only in the germ cells. These studies suggest that the 1.6 kb of Cres 5' flanking sequence contains the required DNA elements for expression in the testis, but lacks the elements to correctly target expression of the reporter gene in the epididymis. Alternatively, repressor elements may be present. Finally, in vitro protease assays were performed to determine if CRES functions as a protease inhibitor. In contrast to cystatin C, CRES did not inhibit the C1 cysteine protease papain but rather inhibited at nanomolar concentrations the serine protease PC2, a prohormone processing enzyme. Therefore, CRES is a new cross class inhibitor that may regulate PC2 or PC2 like proteases and suggests a role for CRES in the regulation of prohormone and proprotein processing.
出处 《中华男科学杂志》 CAS CSCD 2002年第5期313-318,共6页 National Journal of Andrology
基金 NIH NICHDHD3 3 90 3 HoustonEndowment SouthPlainsFoundation资助
关键词 胱蛋白酶抑制剂 附睾 精子发生 基因调控 Cystatin Epididymidis Spermatogenesis Gene regulation
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参考文献19

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同被引文献22

  • 1袁青,徐晨,郭强苏,王一飞.胱蛋白酶抑制剂相关的附睾精子发生基因在小鼠睾丸和附睾中的表达[J].生殖与避孕,2005,25(2):88-88. 被引量:1
  • 2Richardson RT, Sivashanmugam P, Hall SH, et al. Cloning and sequencing of human Eppin: A novel family of protease inhibitors expressed in the epididymis and testis[J]. Gene, 2001, 270(1-2):93-102.
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  • 6Hagiwara K, Kikuchi T, Endo Y, et al. Mouse SWAM1 and SWAM2 are antibacterial proteins composed of a single whey acidic protein motif[J]. J Immunol, 2003, 170(4):1973-1979.
  • 7Yenugu S, Richardson RT, Sivashanmugam P, et al. Antimicrobial activity of human EPPIN, an androgen-regulated, sperm-bound protein with a when acidic protein motif[J]. Biol Reprod, 2004, 71(5):1484-1490.
  • 8Zhou CX, Zhang YL, Xiao L, et al. An epididymis-specific beta-defensin is important for the initiation of sperm maturation[J]. Nat Cell Biol, 2004, 6(5):458-464.
  • 9Ellerman DA, Da Ros VG, Cohen DJ, et al. Expression and structure-function analysis of a sperm cysteine-rich secretory protein that mediates gamete fusion[J]. Biol Reprod, 2002, 67(4):1225-1231.
  • 10O'Rand MG, Widgren EE, Sivashanmugam P, et al. Reversible Immunocontraception in Male Monkeys Immunized with Eppin[J]. Science, 2004, 306(5699):1189-1190.

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