Pennock D G, Thatcher T, Bowen J, Bruns P J, Gorovsky M A
Department of Biology, University of Rochester, New York 14627.
Genetics. 1988 Nov;120(3):697-705. doi: 10.1093/genetics/120.3.697.
Nineteen mutants that are conditional for both the ability to regain motility following deciliation and the ability to grow were isolated. The mutations causing slow growth were placed into five complementation groups. None of the mutations appears to affect energy production as all mutants remained motile at the restrictive temperature. In three complementation groups protein synthesis and the levels of mRNA encoding alpha-tubulin or actin were largely unaffected at the restrictive temperature, consistent with the hypothesis that mutations in these three groups directly affect the assembly of functional cilia and growth. Complementation group 1 was chosen for further characterization. Both phenotypes were shown to be linked, suggesting they are caused by a single mutation. Group 1 mutants regenerated cilia at the restrictive temperature, but the cilia were nonmotile. This mutation also caused a block in cytokinesis at the restrictive temperature but did not affect nuclear divisions or DNA synthesis. The block in cell division was transiently rescued by wild-type cytoplasm exchanged when mutants were paired with wild-type cells during conjugation (round 1 of genomic exclusion). Thus, at least one mutation has been isolated that affects assembly of some microtubule-based structures in Tetrahymena (cilia during regeneration) but not others (nuclei divide at 38 degrees), and the product of this gene is likely to play a role in both ciliary function and in cytokinesis.
分离出了19个突变体,这些突变体在脱纤毛后恢复运动能力以及生长能力方面均表现出条件性。导致生长缓慢的突变被分为五个互补群。由于所有突变体在限制温度下仍保持运动能力,因此似乎没有一个突变影响能量产生。在三个互补群中,蛋白质合成以及编码α-微管蛋白或肌动蛋白的mRNA水平在限制温度下基本未受影响,这与以下假设一致:这三个群中的突变直接影响功能性纤毛的组装和生长。选择互补群1进行进一步表征。两种表型显示是连锁的,表明它们是由单个突变引起的。1组突变体在限制温度下再生纤毛,但这些纤毛无运动能力。该突变在限制温度下还导致胞质分裂受阻,但不影响核分裂或DNA合成。当突变体在接合过程中(基因组排除的第一轮)与野生型细胞配对时,通过交换野生型细胞质可短暂挽救细胞分裂的阻滞。因此,至少分离出了一个影响四膜虫中一些基于微管的结构(再生期间的纤毛)组装但不影响其他结构(细胞核在38摄氏度时分裂)的突变,并且该基因的产物可能在纤毛功能和胞质分裂中均起作用。