Khrebtukova I, Spreitzer R J
Department of Biochemistry, University of Nebraska, Lincoln 68588-0664, USA.
Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13689-93. doi: 10.1073/pnas.93.24.13689.
Ribulose-1,5-bisphosphate carboxylase/ oxygenase (EC 4.1.1.39) is the key photosynthetic enzyme that catalyzes the first step of CO2 fixation. The chloroplast-localized holoenzyme of plants and green algae contains eight nuclear-encoded small subunits and eight chloroplast-encoded large subunits. Although much has been learned about the enzyme active site that resides within each large subunit, it has been difficult to assess the role of eukaryotic small subunits in holoenzyme function and expression. Small subunits are coded by a family of genes, precluding genetic screening or nuclear transformation approaches for the recovery of small-subunit mutants. In this study, the two small-subunit mutants. In this study, the two small-subunit genes of the green alga Chlamydomonas reinhardtii were eliminated during random insertional mutagenesis. The photosynthesis-deficient deletion mutant can be complemented with either of the two wild-type small-subunit genes or with a chimeric gene that contains features of both. Thus, either small subunit is sufficient for holoenzyme assembly and function. In the absence of small subunits, expression of chloroplast-encoded large subunits appears to be inhibited at the level of translation.
1,5 - 二磷酸核酮糖羧化酶/加氧酶(EC 4.1.1.39)是催化二氧化碳固定第一步的关键光合酶。植物和绿藻中定位于叶绿体的全酶包含八个核编码的小亚基和八个叶绿体编码的大亚基。尽管人们对每个大亚基内的酶活性位点已经有了很多了解,但评估真核小亚基在全酶功能和表达中的作用一直很困难。小亚基由一个基因家族编码,这使得通过遗传筛选或核转化方法来获得小亚基突变体变得不可能。在本研究中,绿藻莱茵衣藻的两个小亚基基因在随机插入诱变过程中被消除。光合作用缺陷型缺失突变体可以被两个野生型小亚基基因中的任何一个或包含两者特征的嵌合基因所互补。因此,任何一个小亚基对于全酶组装和功能都是足够的。在没有小亚基的情况下,叶绿体编码的大亚基的表达似乎在翻译水平上受到抑制。