Zhang L, Pakrasi H B, Whitmarsh J
Department of Plant Biology, University of Illinois, Urbana 61801.
J Biol Chem. 1994 Feb 18;269(7):5036-42.
In the cyanobacterium Synechocystis sp. PCC 6803, photosynthetic electron transport from the cytochrome bf complex to photosystem I can be mediated by cytochrome c553 or plastocyanin. The concentration of copper in the growth medium determines which protein is synthesized. To investigate the role of cytochrome c553 in photosynthetic and respiratory electron transport, we cloned the petJ gene encoding cytochrome c553 from Synechocystis 6803 and determined its nucleotide sequence. The 360-base pair open reading frame encodes an 85-amino acid mature protein (predicted molecular mass = 8,742 Da) with a 35-amino acid presequence. Two mutants were constructed, one in which the petJ gene for cytochrome c553 was deleted and another in which the petE gene for plastocyanin was insertionally inactivated. The cytochrome c553 deletion mutant (M109) grew photoautotrophically, even in the absence of copper that prevented the synthesis of plastocyanin, as did the plastocyanin-deficient mutant (M114) grown in the presence of copper that prevented the synthesis of cytochrome c553. The M109 strain exhibited photosynthetic electron transport rates similar to those of wild-type cells when grown under conditions that prevented the synthesis of plastocyanin. Moreover, in M109 cells grown without copper, cytochrome f was completely photooxidized in less than 10 ms by photosystem I. These observations show that electrons can be transferred from the cytochrome bf complex to photosystem I in the absence of both cytochrome c553 and plastocyanin. Additionally, the M109 cells exhibited dark respiration rates comparable with those of wild-type cells, indicating that cytochrome c553 is not obligately required for respiratory electron transport in Synechocystis 6803.
在蓝藻集胞藻PCC 6803中,从细胞色素bf复合物到光系统I的光合电子传递可由细胞色素c553或质体蓝素介导。生长培养基中铜的浓度决定合成哪种蛋白质。为了研究细胞色素c553在光合和呼吸电子传递中的作用,我们从集胞藻6803中克隆了编码细胞色素c553的petJ基因,并测定了其核苷酸序列。360个碱基对的开放阅读框编码一个含35个氨基酸前导序列的85个氨基酸的成熟蛋白(预测分子量 = 8742道尔顿)。构建了两个突变体,一个是细胞色素c553的petJ基因缺失的突变体,另一个是质体蓝素的petE基因插入失活的突变体。细胞色素c553缺失突变体(M109)能够光自养生长,即使在缺乏阻止质体蓝素合成的铜的情况下也是如此,在存在阻止细胞色素c553合成的铜的情况下生长的质体蓝素缺陷突变体(M114)也是如此。当在阻止质体蓝素合成的条件下生长时,M109菌株表现出与野生型细胞相似的光合电子传递速率。此外,在无铜条件下生长的M109细胞中,细胞色素f在不到10毫秒的时间内被光系统I完全光氧化。这些观察结果表明,在细胞色素c553和质体蓝素都不存在的情况下,电子可以从细胞色素bf复合物转移到光系统I。此外,M109细胞表现出与野生型细胞相当的暗呼吸速率,表明细胞色素c553不是集胞藻6803呼吸电子传递所必需的。