Gu W, Wu X Q, Meng X H, Morales C, el-Alfy M, Hecht N B
Department of Biology, Tufts University, Medford, Massachusetts, USA.
Mol Reprod Dev. 1998 Mar;49(3):219-28. doi: 10.1002/(SICI)1098-2795(199803)49:3<219::AID-MRD1>3.0.CO;2-R.
Testis brain RNA-binding protein (TB-RBP) suppresses translation in vitro and attaches mRNAs to microtubules by binding to conserved elements in the 3' untranslated regions (UTRs) of specific testis and brain mRNAs. Purification of TB-RBP from testicular and brain cytoplasmic extracts has revealed that mouse TB-RBP is 99% identical to the human protein translin, a recombination "hot spot" binding protein associated with chromosomal translocations. Using a cDNA encoding TB-RBP, the gene copy number and the developmental expression of TB-RBP have been analyzed by Southern blotting, Northern blotting, and in situ hybridization. In the mouse, TB-RBP is encoded by a single copy gene. In mouse testes, three TB-RBP mRNAs of about 1.2, 1.7, and 3.0 kb are developmentally regulated with high levels of expression in meiotic and postmeiotic germ cells. A fourth TB-RBP transcript of about 3.2 kb is seen in the brain. In situ hybridization confirms high levels of testicular TB-RBP mRNAs in meiotic and postmeiotic cells, with the highest levels of TB-RBP mRNAs in pachytene spermatocytes and round spermatids of the mouse and in round spermatids of the rat. RNase H digestion assays reveal that the three TB-RBP mRNAs of mouse testes result from processing differences in their 3' untranslated regions. These data demonstrate that multiple TB-RBP mRNAs are primarily expressed in meiotic and postmeiotic germ cells in the mammalian testis, and although the specific RNA-binding ability of TB-RBP appears limited to brain and testis, TB-RBP mRNAs are widely expressed.
睾丸脑RNA结合蛋白(TB-RBP)在体外抑制翻译,并通过与特定睾丸和脑mRNA的3'非翻译区(UTR)中的保守元件结合,将mRNA附着于微管。从睾丸和脑细胞质提取物中纯化TB-RBP表明,小鼠TB-RBP与人蛋白转座蛋白99%相同,转座蛋白是一种与染色体易位相关的重组“热点”结合蛋白。利用编码TB-RBP的cDNA,通过Southern印迹、Northern印迹和原位杂交分析了TB-RBP的基因拷贝数和发育表达情况。在小鼠中,TB-RBP由单拷贝基因编码。在小鼠睾丸中,约1.2、1.7和3.0 kb的三种TB-RBP mRNA在减数分裂和减数分裂后生殖细胞中受到发育调控,在减数分裂和减数分裂后生殖细胞中表达水平较高。在脑中可见约3.2 kb的第四种TB-RBP转录本。原位杂交证实减数分裂和减数分裂后细胞中睾丸TB-RBP mRNA水平较高,在小鼠的粗线期精母细胞和圆形精子细胞以及大鼠的圆形精子细胞中TB-RBP mRNA水平最高。RNase H消化试验表明,小鼠睾丸中的三种TB-RBP mRNA是由其3'非翻译区的加工差异产生的。这些数据表明,多种TB-RBP mRNA主要在哺乳动物睾丸的减数分裂和减数分裂后生殖细胞中表达,尽管TB-RBP的特异性RNA结合能力似乎仅限于脑和睾丸,但TB-RBP mRNA广泛表达。