Cold Spring Harbor Laboratory, P.O. Box 100, Cold Spring Harbor, NY 11724.
Proc Natl Acad Sci U S A. 1986 Apr;83(7):2012-6. doi: 10.1073/pnas.83.7.2012.
The structural gene for cytochrome c(2) (cycA) of the photosynthetic bacterium Rhodopseudomonas capsulata has been cloned, and the nucleotide and deduced polypeptide sequences have been determined. Compared with the known amino acid sequence of the purified cytochrome c(2), the nucleotide sequence corresponding to the N-terminal part of the cycA gene product indicates the presence of a putative 21 amino acid signal sequence. Thus, cytochrome c(2) may be synthesized as a precursor which is processed during its secretion to the periplasm. Insertion and insertion-deletion mutations were constructed in vitro and the chromosomal cycA(+) allele of a wild-type strain was replaced with these mutations by homologous recombination to yield c(2) (-) mutants of R. capsulata. The c(2) (-) mutants are stable, and they can grow by photosynthesis and by respiration. Since cytochrome c(2) is the primary electron donor to the reaction center during photosynthesis, the ability of these mutants to grow photosynthetically indicates that an alternative way(s) of reducing the oxidized reaction center must exist in R. capsulata. One candidate for this role may be the membrane-bound cytochrome c(1).
已克隆了光合细菌荚膜红假单胞菌细胞色素 c(2)(cycA)的结构基因,并测定了其核苷酸和推导的多肽序列。与已纯化的细胞色素 c(2)的已知氨基酸序列相比,cycA 基因产物的 N 末端部分的核苷酸序列表明存在一个推定的 21 个氨基酸信号序列。因此,细胞色素 c(2)可能作为前体合成,在其分泌到周质期间进行加工。体外构建了插入和插入缺失突变,并通过同源重组用这些突变取代野生型菌株的染色体 cycA(+)等位基因,得到荚膜红假单胞菌的 c(2)(-)突变体。c(2)(-)突变体稳定,它们可以通过光合作用和呼吸作用生长。由于细胞色素 c(2)是光合作用期间向反应中心的主要电子供体,这些突变体进行光合作用的能力表明荚膜红假单胞菌中必须存在替代的还原氧化反应中心的方式。这种作用的一个候选者可能是膜结合的细胞色素 c(1)。