Rasooly R S, New C M, Zhang P, Hawley R S, Baker B S
Department of Molecular Genetics, Albert Einstein College of Medicine, Bronx, New York 10461.
Genetics. 1991 Oct;129(2):409-22. doi: 10.1093/genetics/129.2.409.
The l(1)TW-6cs mutation is a cold-sensitive recessive lethal mutation in Drosophila melanogaster, that affects both meiotic and mitotic chromosome segregation. We report the isolation of three revertants of this mutation. All three revert both the meiotic and mitotic effects as well as the cold sensitivity, demonstrating that all three phenotypes are due to a single lesion. We further show that these revertants fail to complement an amorphic allele of the nod (no distributive disjunction) locus, which encodes a kinesin-like protein. These experiments demonstrate that l(1)TW-6cs is an antimorphic allele of nod, and we rename it nodDTW. Sequencing of the nod locus on a nodDTW-bearing chromosome reveals a single base change in the putative ATP-binding region of the motor domain of nod. Recessive, loss-of-function mutations at the nod locus specifically disrupt the segregation of nonexchange chromosomes in female meiosis. We demonstrate that, at 23.5 degrees, the meiotic defects in nodDTW/+ females are similar to those observed in nod/nod females; that is, the segregation of nonexchange chromosomes is abnormal. However, in nodDTW/nodDTW females, or in nodDTW/+ females at 18 degrees, we observe a more severe meiotic defect that apparently affects the segregation of both exchange and nonexchange chromosomes. In addition, nodDTW homozygotes and hemizygous males have previously been shown to exhibit mitotic defects including somatic chromosome breakage and loss. We propose that the defective protein encoded by the nodDTW allele interferes with proper chromosome movement during both meiosis and mitosis, perhaps by binding irreversibly to microtubules.
l(1)TW - 6cs突变是黑腹果蝇中的一种冷敏感隐性致死突变,它会影响减数分裂和有丝分裂中的染色体分离。我们报告了该突变的三个回复突变体的分离情况。所有这三个回复突变体都恢复了减数分裂和有丝分裂效应以及冷敏感性,这表明所有这三种表型都是由单个损伤引起的。我们进一步表明,这些回复突变体不能与nod(无分布性分离)基因座的一个无效等位基因互补,该基因座编码一种驱动蛋白样蛋白。这些实验证明l(1)TW - 6cs是nod的一个反效等位基因,我们将其重新命名为nodDTW。在携带nodDTW的染色体上对nod基因座进行测序,发现在nod运动结构域的假定ATP结合区域有一个单碱基变化。nod基因座上的隐性功能丧失突变会特异性地破坏雌性减数分裂中非交换染色体的分离。我们证明,在23.5摄氏度时,nodDTW/+雌性中的减数分裂缺陷与在nod/nod雌性中观察到的缺陷相似;也就是说,非交换染色体的分离是异常的。然而,在nodDTW/nodDTW雌性中,或者在18摄氏度的nodDTW/+雌性中,我们观察到更严重的减数分裂缺陷,显然影响了交换染色体和非交换染色体的分离。此外,之前已经表明nodDTW纯合子和半合子雄性表现出有丝分裂缺陷,包括体细胞染色体断裂和丢失。我们提出,由nodDTW等位基因编码的缺陷蛋白可能通过不可逆地结合微管,在减数分裂和有丝分裂过程中干扰染色体的正常移动。