Okkema P G, Kimble J
Department of Biochemistry, College of Agricultural and Life Sciences, Graduate School, University of Wisconsin-Madison 53706.
EMBO J. 1991 Jan;10(1):171-6. doi: 10.1002/j.1460-2075.1991.tb07933.x.
We have cloned the Caenorhabditis elegans sex determining gene, tra-2, by transposon tagging. The tra-2 region is delineated by mapping Southern blot differences associated with 11 tra-2 mutations, mutant rescue, and analysis of tra-2 RNAs. The tra-2 gene encodes three transcripts. One transcript, a 1.8 kb RNA, is not detected in animals lacking a germ line, and therefore may be germline specific. Comparison of the two sexes shows that adult hermaphrodites have approximately 15-fold more tra-2 RNA than adult males. In addition, adult hermaphrodites contain 5 kb and 1.8 kb tra-2 RNAs whereas adult males possess 5 kb and 1.9 kb RNAs. A 1.9 kb tra-2 RNA is also found during hermaphrodite larval development, prior to sexual differentiation of the XX germ line. Surprisingly, analysis of tra-2 expression in selected sex determination mutants reveals that the sex specificity of tra-2 RNAs is not dictated by the pathway of sex determination that has been established by genetic experiments. This result can be interpreted in two ways. Either the sex specificity of the tra-2 RNAs is irrelevant to regulation of the sexual phenotype, which seems unlikely, or there is additional complexity within the hierarchy of sex determining genes, such as feedback regulation, which ensures that the tra-2 product corresponds to phenotypic sex.
我们通过转座子标签法克隆了秀丽隐杆线虫的性别决定基因tra-2。通过对与11个tra-2突变相关的Southern印迹差异进行定位、突变体拯救以及对tra-2 RNA的分析,确定了tra-2区域。tra-2基因编码三种转录本。其中一种转录本,即1.8 kb的RNA,在缺乏生殖系的动物中未被检测到,因此可能是生殖系特异性的。对两性的比较表明,成年雌雄同体的tra-2 RNA比成年雄性大约多15倍。此外,成年雌雄同体含有5 kb和1.8 kb的tra-2 RNA,而成年雄性则拥有5 kb和1.9 kb的RNA。在XX生殖系性别分化之前的雌雄同体幼虫发育过程中也发现了1.9 kb的tra-2 RNA。令人惊讶的是,对选定的性别决定突变体中tra-2表达的分析表明,tra-2 RNA的性别特异性并非由遗传实验所确立的性别决定途径所决定。这个结果可以有两种解释。要么tra-2 RNA的性别特异性与性表型的调控无关,这似乎不太可能,要么在性别决定基因层次结构中存在额外的复杂性,比如反馈调节,以确保tra-2产物与表型性别相对应。