Shibata M, Ohkawa H, Kaneko T, Fukuzawa H, Tabata S, Kaplan A, Ogawa T
Bioscience Center, Nagoya University, Chikusa, Nagoya 464-8601, Japan.
Proc Natl Acad Sci U S A. 2001 Sep 25;98(20):11789-94. doi: 10.1073/pnas.191258298. Epub 2001 Sep 18.
Cyanobacteria possess a CO(2)-concentrating mechanism that involves active CO(2) uptake and HCO(3)(-) transport. For CO(2) uptake, we have identified two systems in the cyanobacterium Synechocystis sp. strain PCC 6803, one induced at low CO(2) and one constitutive. The low CO(2)-induced system showed higher maximal activity and higher affinity for CO(2) than the constitutive system. On the basis of speculation that separate NAD(P)H dehydrogenase complexes were essential for each of these systems, we reasoned that inactivation of one system would allow selection of mutants defective in the other. Thus, mutants unable to grow at pH 7.0 in air were recovered after transformation of a DeltandhD3 mutant with a transposon-bearing library. Four of them had tags within slr1302 (designated cupB), a homologue of sll1734 (cupA), which is cotranscribed with ndhF3 and ndhD3. The DeltacupB, DeltandhD4, and DeltandhF4 mutants showed CO(2)-uptake characteristics of the low CO(2)induced system observed in wild type. In contrast, mutants DeltacupA, DeltandhD3, and DeltandhF3 showed characteristics of the constitutive CO(2)-uptake system. Double mutants impaired in one component of each of the systems were unable to take up CO(2) and required high CO(2) for growth. Phylogenetic analysis indicated that the ndhD3/ndhD4-, ndhF3/ndhF4-, and cupA/cupB-type genes are present only in cyanobacteria. Most of the cyanobacterial strains studied possess the ndhD3/ndhD4-, ndhF3/ndhF4-, and cupA/cupB-type genes in pairs. Thus, the two types of NAD(P)H dehydrogenase complexes essential for low CO(2)-induced and constitutive CO(2)-uptake systems associated with the NdhD3/NdhF3/CupA-homologues and NdhD4/NdhF4/CupB-homologues, respectively, appear to be present in these cyanobacterial strains but not in other organisms.
蓝细菌拥有一种二氧化碳浓缩机制,该机制涉及主动摄取二氧化碳和转运碳酸氢根离子。对于二氧化碳摄取,我们在集胞藻属PCC 6803蓝细菌中鉴定出了两个系统,一个在低二氧化碳条件下诱导产生,另一个为组成型。低二氧化碳诱导系统比组成型系统表现出更高的最大活性和对二氧化碳的更高亲和力。基于推测认为不同的NAD(P)H脱氢酶复合物对于这些系统中的每一个都是必不可少的,我们推断使一个系统失活将允许筛选出在另一个系统中有缺陷的突变体。因此,在用携带转座子的文库转化ΔndhD3突变体后,获得了在空气中pH 7.0条件下无法生长的突变体。其中四个在slr1302(命名为cupB)中有标签,slr1302是sll1734(cupA)的同源物,sll1734与ndhF3和ndhD3共转录。ΔcupB、ΔndhD4和ΔndhF4突变体表现出野生型中观察到的低二氧化碳诱导系统的二氧化碳摄取特征。相反,ΔcupA、ΔndhD3和ΔndhF3突变体表现出组成型二氧化碳摄取系统的特征。在每个系统的一个组分中受损的双突变体无法摄取二氧化碳,并且需要高浓度二氧化碳才能生长。系统发育分析表明,ndhD3/ndhD4、ndhF3/ndhF4和cupA/cupB型基因仅存在于蓝细菌中。大多数研究的蓝细菌菌株成对拥有ndhD3/ndhD4、ndhF3/ndhF4和cupA/cupB型基因。因此,分别与NdhD3/NdhF3/CupA同源物和NdhD4/NdhF4/CupB同源物相关的、对于低二氧化碳诱导和组成型二氧化碳摄取系统必不可少的两种类型的NAD(P)H脱氢酶复合物似乎存在于这些蓝细菌菌株中,而不存在于其他生物体中。