Levitan D J, Boyd L, Mello C C, Kemphues K J, Stinchcomb D T
Department of Cellular and Developmental Biology, Harvard University, Cambridge, MA 02138.
Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):6108-12. doi: 10.1073/pnas.91.13.6108.
The par-2 gene of Caenorhabditis elegans functions in early embryogenesis to ensure an asymmetric first cleavage and the segregation of cytoplasmic factors. Both processes appear to be required to generate daughter blastomeres with distinct developmental potential. We isolated an allele of par-2 by using a screen for maternal-effect lethal mutations in a strain known for its high frequency of transposition events. A transposable element was found to be linked to this allele. Sequences flanking the site of transposon insertion were cloned and found to rescue the par-2 mutant phenotype. DNA in the par-2 region hybridized to a 2.3-kb germ-line-enriched mRNA. The cDNA corresponding to this germ-line-enriched message was cloned, sequenced, and used to identify the molecular lesions associated with three par-2 alleles. Sequence analysis of the par-2 cDNA revealed that the predicted protein contained two distinct motifs found in other known proteins: an ATP-binding site and a cysteine-rich motif which identifies the par-2 gene product as a member of a growing class of putative zinc-binding proteins.
秀丽隐杆线虫的par-2基因在早期胚胎发生过程中发挥作用,以确保第一次分裂的不对称性以及细胞质因子的分离。这两个过程似乎都是产生具有不同发育潜能的子细胞所必需的。我们通过在一个以转座事件高频发生而闻名的品系中筛选母性效应致死突变,分离出了一个par-2等位基因。发现一个转座元件与该等位基因连锁。克隆了转座子插入位点两侧的序列,并发现其可挽救par-2突变体表型。par-2区域的DNA与一个2.3 kb的生殖系富集mRNA杂交。克隆了与这种生殖系富集信息相对应的cDNA,进行了测序,并用于鉴定与三个par-2等位基因相关的分子损伤。par-2 cDNA的序列分析表明,预测的蛋白质包含在其他已知蛋白质中发现的两个不同基序:一个ATP结合位点和一个富含半胱氨酸的基序,该基序将par-2基因产物鉴定为一类不断增加的假定锌结合蛋白的成员。