IDENTIFICATION OF THE GENE FOR THE DEVELOPMENTALLY EXPRESSED 70KDA HEAT-SHOCK PROTEIN (P70) OF MOUSE SPERMATOGENIC CELLS
IDENTIFICATION OF THE GENE FOR THE DEVELOPMENTALLY EXPRESSED 70KDA HEAT-SHOCK PROTEIN (P70) OF MOUSE SPERMATOGENIC CELLS
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DOI：
10.1016/0012-1606(92)90002-x
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发表时间：
1992-03-01
影响因子：
2.7
通讯作者：
EDDY, EM
中科院分区：
文献类型：
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作者：
ROSARIO, MO;PERKINS, SL;EDDY, EM
小鼠生精细胞合成与哺乳动物细胞的主要热休克蛋白 (hsp70) 密切相关的 70 kDa 蛋白 (P70)（R. L. Allen, D. A. O'Brien, and E. M. Eddy,Mol. Cell. Biol.8,828–832, 1988）。 P70 的表达在发育过程中受到调节，而 hsp70 则在应激反应中被诱导，这表明 P70 是在生精细胞中转录的 Hsp70 多基因家族的独特成员的产物。该基因的有力候选者是 Hsp70.2 基因（Z. F. Zakeri、D. J. Wolgemuth 和 C. R. Hunt, Mol. Cell. Biol.8,2925–2932, 1988）。来自 Hsp70.2 5' 区域的 DNA 片段与 2.7 kb 转录物杂交，在小鼠生精细胞中的表达时间模式与 P70 蛋白相似。我们使用针对从Hsp70.2序列预测的合成肽产生的多克隆抗血清来表征其蛋白质产物并从粗线期精母细胞表达文库中分离cDNA克隆。抗血清与小鼠睾丸切片上的减数分裂和减数分裂后生精细胞发生特异性反应。它识别二维凝胶蛋白质印迹上的 P70 蛋白，并且不与生精细胞或体细胞的其他热休克蛋白结合。 cDNA 与 2.7 kb mRNA 杂交，该 mRNA 在未受应激的粗线期精母细胞和圆形精子细胞中含量丰富，但在其他细胞类型中未检测到。对两个 cDNA 进行测序，发现与 Hsp70.2 基因的 3' 端具有 99% 同源性。这些数据有力地支持了以下假设：P70是小鼠生精细胞中Hsp70.2基因的表达产物。
Mouse spermatogenic cells synthesize a 70-kDa protein (P70) closely related to the major heat-shock protein (hsp70) of mammalian cells (R. L. Allen, D. A. O'Brien, and E. M. Eddy,Mol. Cell. Biol.8,828–832, 1988). Expression of P70 is developmentally regulated while hsp70 is induced in response to stress, suggesting that P70 is the product of a unique member of theHsp70multigene family transcribed in spermatogenic cells. A strong candidate for this gene was theHsp70.2gene (Z. F. Zakeri, D. J. Wolgemuth, and C. R. Hunt,Mol. Cell. Biol.8,2925–2932, 1988). A DNA segment from the 5′ region ofHsp70.2hybridized to a 2.7-kb transcript with a temporal pattern of expression in mouse spermatogenic cells similar to the P70 protein. We used a polyclonal antiserum generated against a synthetic peptide predicted from theHsp70.2sequence to characterize its protein product and to isolate cDNA clones from a pachytene spermatocyte expression library. The antiserum reacted specifically with meiotic and postmeiotic spermatogenic cells on sections of mouse testis. It recognized the P70 protein on Western blots of two-dimensional gels and did not bind to other heat-shock proteins of spermatogenic or somatic cells. The cDNAs hybridized to a 2.7-kb mRNA that was abundant in unstressed pachytene spermatocytes and round spermatids but was not detected in other cell types. Two cDNAs were sequenced and found to be 99% homologous to the 3′ end of theHsp70.2gene. These data strongly supported the hypothesis that P70 is the expressed product of theHsp70.2gene in mouse spermatogenic cells.