Brown Jason M, Mosley Matthew, Montes-Berrueta Daniela, Hou Yuqing, Yang Fan, Scarbrough Chasity, Witman George B, Wirschell Maureen
Department of Biology, Salem State University, Salem, Massachusetts, United States of America.
University of Mississippi Medical Center, Department of Biochemistry, Jackson, Mississippi, United States of America.
PLoS One. 2017 Mar 14;12(3):e0173842. doi: 10.1371/journal.pone.0173842. eCollection 2017.
We have used an insertional mutagenesis approach to generate new C. reinhardtii motility mutants. Of 56 mutants isolated, one is a new allele at the ODA3 locus, called oda3-6. Similar to the previously characterized oda3 alleles, oda3-6 has a slow-jerky swimming phenotype and reduced swimming speed. The oda3-6 mutant fails to assemble the outer dynein arm motor and outer dynein arm-docking complex (ODA-DC) in the ciliary axoneme due to an insertion in the 5' end of the DCC1 gene, which encodes the DC1 subunit of the ODA-DC. Transformation of oda3-6 with the wild-type DCC1 gene rescues the mutant swimming phenotype and restores assembly of the ODA-DC and the outer dynein arm in the cilium. This is the first oda3 mutant to be characterized at the molecular level and is likely to be very useful for further analysis of DC1 function.
我们采用插入诱变方法来生成新的莱茵衣藻运动突变体。在分离出的56个突变体中,有一个是ODA3基因座的新等位基因,称为oda3-6。与先前鉴定的oda3等位基因相似,oda3-6具有缓慢且不连贯的游动表型,游动速度降低。由于DCC1基因5'端的插入,oda3-6突变体无法在纤毛轴丝中组装外动力蛋白臂马达和外动力蛋白臂对接复合体(ODA-DC),该基因编码ODA-DC的DC1亚基。用野生型DCC1基因转化oda3-6可挽救突变体的游动表型,并恢复纤毛中ODA-DC和外动力蛋白臂的组装。这是第一个在分子水平上得到表征的oda3突变体,可能对进一步分析DC1功能非常有用。