Yang J, Porter L, Rawls J
Molecular Cell Biology Group, T.H. Morgan School of Biological Sciences, University of Kentucky, Lexington 40506.
Mol Gen Genet. 1995 Feb 6;246(3):334-41. doi: 10.1007/BF00288606.
The dhod gene encodes dihydroorotate dehydrogenase (DHOdehase), which catalyzes the fourth step of de novo pyrimidine biosynthesis. In addition to the common 1.5 kb dhod RNA expressed by embryos and females, adult males produce a group of slightly longer RNAs. Evidence is presented that the latter RNAs arise through transcription initiation at sites upstream from that of the common RNA and expression of these male-specific RNAs is limited to spermatogenesis. In situ hybridization analysis shows that these RNAs accumulate during spermatocyte growth and persist through meiosis and early spermatid differentiation. In contrast, DHOdehase activity is virtually absent in spermatocytes, meiotic cells, and in early spermatid cysts, then it becomes highly abundant in elongated spermatid cysts and disappears in late spermatogenesis. Thus, testis-limited expression of dhod conforms to a model proposed for other genes that function during spermiogenesis: transcription in spermatocytes, storage of translationally inactive RNA through meiosis, translation of the RNA during spermiogenesis. Very similar expression of a testis promoter-lacZ fusion transgene indicates that sequences required for the spermatogenesis transcription and translation patterns are confined to the 5' end of the dhod gene. Deletion analysis of that 5' region delimits all sequences necessary for spermatid expression of the transgene to a 89 bp fragment. These results are discussed in the contexts of known mechanisms of gene regulation during spermatogenesis and potential roles of DHOdehase during spermiogenesis.
dhod基因编码二氢乳清酸脱氢酶(DHO脱氢酶),该酶催化嘧啶从头生物合成的第四步。除了胚胎和雌性表达的常见1.5 kb dhod RNA外,成年雄性还产生一组略长的RNA。有证据表明,后者的RNA是通过在常见RNA上游位点的转录起始产生的,并且这些雄性特异性RNA的表达仅限于精子发生过程。原位杂交分析表明,这些RNA在精母细胞生长过程中积累,并在减数分裂和早期精子细胞分化过程中持续存在。相比之下,DHO脱氢酶活性在精母细胞、减数分裂细胞和早期精子细胞囊肿中几乎不存在,然后在伸长的精子细胞囊肿中变得高度丰富,并在精子发生后期消失。因此,dhod在睾丸中的局限性表达符合为其他在精子形成过程中起作用的基因提出的模型:在精母细胞中进行转录,通过减数分裂储存翻译无活性的RNA,在精子形成过程中进行RNA的翻译。睾丸启动子-lacZ融合转基因的非常相似的表达表明,精子发生转录和翻译模式所需的序列局限于dhod基因的5'端。对该5'区域的缺失分析将转基因在精子细胞中表达所需的所有序列限定为一个89 bp的片段。在精子发生过程中已知的基因调控机制以及DHO脱氢酶在精子形成过程中的潜在作用的背景下讨论了这些结果。