O'Donnell K L, Osmani A H, Osmani S A, Morris N R
Department of Pharmacology, University of Medicine and Dentistry of New Jersey, Piscataway 08854.
J Cell Sci. 1991 Aug;99 ( Pt 4):711-9. doi: 10.1242/jcs.99.4.711.
The recessive, temperature-sensitive bimA1 mutation of Aspergillus nidulans blocks nuclei in metaphase at restrictive temperature. To determine whether the bimA product is essential, integrative transformation was used to create a mutation in the bimA gene. The mutation was maintained in a heterokaryon and the phenotype of spores produced by the heterokaryon was analyzed. Molecular disruption of the wild-type bimA gene is recessive in the heterokaryon and causes a metaphase block, demonstrating that bimA is an essential gene for mitosis. bimA was cloned by DNA-mediated complementation of its mutant phenotype at restrictive temperature, and the nucleotide sequence of a full-length cDNA was determined. A single large open reading frame was identified in the cDNA sequence, which predicts a protein containing 806 amino acid residues that is related (30.4% identity) to the Schizosaccharomyces pombe nuc2+ gene product, which also is required for completion of mitosis. The sequence of the bimA gene indicates that it is a member of a family of mostly nuclear proteins that contain a degenerate 34 amino acid repeat, the TPR (tetratricopeptide repeat) gene family.
构巢曲霉的隐性温度敏感型bimA1突变在限制温度下使细胞核阻滞于中期。为确定bimA产物是否必不可少,采用整合转化法在bimA基因中制造突变。该突变在异核体中得以维持,并对异核体产生的孢子的表型进行了分析。野生型bimA基因的分子破坏在异核体中呈隐性,并导致中期阻滞,这表明bimA是有丝分裂的必需基因。通过在限制温度下对其突变表型进行DNA介导的互补作用克隆了bimA,并测定了全长cDNA的核苷酸序列。在cDNA序列中鉴定出一个单一的大开放阅读框,其预测的蛋白质含有806个氨基酸残基,与粟酒裂殖酵母nuc2+基因产物相关(同一性为30.4%),该基因产物对于完成有丝分裂也是必需的。bimA基因的序列表明它是一个主要由核蛋白组成的家族的成员,该家族包含一个简并的34个氨基酸重复序列,即TPR(四肽重复)基因家族。