Pérez-Martínez X, Vazquez-Acevedo M, Tolkunova E, Funes S, Claros M G, Davidson E, King M P, González-Halphen D
Departamento de Genética Molecular, Instituto de Fisiologia Celular, Universidad Nacional Autónoma de México, 04510.
J Biol Chem. 2000 Sep 29;275(39):30144-52. doi: 10.1074/jbc.M003940200.
The algae of the family Chlamydomonadaceae lack the gene cox3 that encodes subunit III of cytochrome c oxidase in their mitochondrial genomes. This observation has raised the question of whether this subunit is present in cytochrome c oxidase or whether the corresponding gene is located in the nucleus. Cytochrome c oxidase was isolated from the colorless chlamydomonad Polytomella spp., and the existence of subunit III was established by immunoblotting analysis with an antibody directed against Saccharomyces cerevisiae subunit III. Based partly upon the N-terminal sequence of this subunit, oligodeoxynucleotides were designed and used for polymerase chain reaction amplification, and the resulting product was used to screen a cDNA library of Chlamydomonas reinhardtii. The complete sequences of the cox3 cDNAs from Polytomella spp. and C. reinhardtii are reported. Evidence is provided that the genes for cox3 are encoded by nuclear DNA, and the predicted polypeptides exhibit diminished physical constraints for import as compared with mitochondrial-DNA encoded homologs. This indicates that transfer of this gene to the nucleus occurred before Polytomella diverged from the photosynthetic Chlamydomonas lineage and that this transfer may have occurred in all chlamydomonad algae.
衣藻科藻类的线粒体基因组中缺乏编码细胞色素c氧化酶亚基III的基因cox3。这一发现引发了一个问题:该亚基是否存在于细胞色素c氧化酶中,或者相应的基因是否位于细胞核中。从无色衣藻多鞭藻属(Polytomella spp.)中分离出细胞色素c氧化酶,并用针对酿酒酵母亚基III的抗体通过免疫印迹分析确定了亚基III的存在。部分基于该亚基的N端序列设计了寡脱氧核苷酸,并用于聚合酶链反应扩增,所得产物用于筛选莱茵衣藻(Chlamydomonas reinhardtii)的cDNA文库。报道了多鞭藻属和莱茵衣藻cox3 cDNA的完整序列。有证据表明,cox3基因由核DNA编码,与线粒体DNA编码的同源物相比,预测的多肽在导入时受到的物理限制较小。这表明该基因转移到细胞核发生在多鞭藻从光合衣藻谱系分化之前,并且这种转移可能发生在所有衣藻藻类中。