Carvalho A B, Dobo B A, Vibranovski M D, Clark A G
Departamento de Genética, Universidade Federal do Rio de Janeiro, Caixa Postal 68011 CEP 21944-970, Rio de Janeiro, Brazil.
Proc Natl Acad Sci U S A. 2001 Nov 6;98(23):13225-30. doi: 10.1073/pnas.231484998. Epub 2001 Oct 30.
The heterochromatic state of the Drosophila Y chromosome has made the cloning and identification of Y-linked genes a challenging process. Here, we report application of a procedure to identify Y-linked gene fragments from the unmapped residue of the whole genome sequencing effort. Previously identified Y-linked genes appear in sequenced scaffolds as individual exons, apparently because many introns have become heterochromatic, growing to enormous size and becoming virtually unclonable. A TBLASTN search using all known proteins as query sequences, tested against a blastable database of the unmapped fragments, produced a number of matches consistent with this scenario. Reverse transcription-PCR and genetic methods were used to confirm those that are expressed, Y-linked genes. The five genes reported here include three protein phosphatases (Pp1-Y1, Pp1-Y2, and PPr-Y), an occludin-related gene (ORY), and a coiled-coils gene (CCY). This brings the total to nine protein-coding genes identified on the Drosophila Y chromosome. ORY and CCY may correspond, respectively, to the fertility factors ks-1 and ks-2, whereas the three protein phosphatases represent novel genes. There remains a strong functional coherence to male function among the genes on the Drosophila Y chromosome.
果蝇Y染色体的异染色质状态使得Y连锁基因的克隆和鉴定成为一个具有挑战性的过程。在这里,我们报告了一种从全基因组测序工作的未定位残基中鉴定Y连锁基因片段的方法的应用。先前鉴定的Y连锁基因在测序支架中以单个外显子的形式出现,显然是因为许多内含子已经变成异染色质,变得非常大,几乎无法克隆。使用所有已知蛋白质作为查询序列进行TBLASTN搜索,并针对未定位片段的可比对数据库进行测试,得到了一些与这种情况相符的匹配结果。使用逆转录PCR和遗传学方法来确认那些表达的Y连锁基因。这里报道的五个基因包括三个蛋白磷酸酶(Pp1-Y1、Pp1-Y2和PPr-Y)、一个闭合蛋白相关基因(ORY)和一个卷曲螺旋基因(CCY)。这使得在果蝇Y染色体上鉴定出的蛋白质编码基因总数达到九个。ORY和CCY可能分别对应于育性因子ks-1和ks-2,而这三个蛋白磷酸酶代表新基因。果蝇Y染色体上的基因之间在雄性功能方面仍然存在很强的功能一致性。