Cloning and transcription analysis of the ndh(A-I-G-E) gene cluster and the ndhD gene of the cyanobacterium Synechocystis sp. PCC6803

Abstract

The plastid DNA of higher plants contains eleven reading frames that are homologous to subunits of the mitochondrial NADH-ubiquinone oxidoreductase (complex I). The genes are expressed, but a plastid NAD(P)H dehydrogenase has not yet been isolated and the function of the enzyme in plastid metabolism is unknown. Cyanobacteria also contain a NADH dehydrogenase that is homologous to the mitochondrial complex I. The enzyme is sensitive to rotenone and is located on the cytoplasmic and the thylakoid membrane. We report here the sequence of five subunits (ndhA, -I, G, -E and -D) of the NADH dehydrogenase from the unicellular cyanobacterium Synechocystis sp. PCC6803. As in plastid DNA, the genes ndh(A-I-G-E) are clustered and probably constitute an operon. The ndhD gene is associated with a gene encoding an iron-sulphur protein of photosystem I (psaC) as in plastid DNA. In contrast to the situation in plastids, psaC and ndhD are not cotranscribed but transcribed from opposite strands. The deduced amino acid sequence of the cyanobacterial polypeptides is more similar to the corresponding plastid (40-68% identity) than to the corresponding mitochondrial subunits (17-39% identity). Thus, the cyanobacterial NADH-dehydrogenase provides a prokaryotic model system which is more suitable to genetic analysis than the enzyme of plastids.

Immunopurification of a subcomplex of the NAD(P)H‐plastoquinone‐oxidoreductase from the cyanobacterium Synechocystis sp. PCC6803

“…The native NDH-H subunit lacks the aminoterminal methionine. The sequences of the corresponding genes have been described recently [7,23].…”

“…PCC6803 are known [6,7,23,27,28]. Among these genes are seven subunits (ndhA-G) of which homologue (ND1-ND4L-ND6) are encoded in the mitochondrial genome of vertebrates [3].…”

“…The ndhF gene has also been isolated and partially sequenced (E1-lersiek and Steinmtiller, in preparation). However, no corresponding proteins to these genes were found in the Table I Protein sequences of the subunits of the purified subcomplex The sequences from the amino-terminal amino acid analysis are compared to the sequences deduced from the DNA sequence of the corresponding genes published in [6,7,23]. From the 21 kDa band two sequences were obtained: a dominant sequence (al) and a side sequence (a2).…”

Genome-Wide Dynamic Transcriptional Profiling of the Light-to-Dark Transition inSynechocystissp. Strain PCC 6803

“…Specifically, ndbB (slr1743; 3 total ndb genes [31]), ndhD (slr2007; 5 ndhD genes [50]), ndhD2 (slr1291; 5 genes [50]), ndhF1 (sll1732; 4 genes), and ndhF2 (slr0844; 4 genes) transcripts all increased, while solitary genes, such as ndhA, ndhB, ndhC, and ndhE, did not show transcript accumulation in the presence of light. Also, expression from the operon ndhAIGE (18) and gene clusters ndhCKJ and ndhFD were internally consistent and did not change from dark levels (data not shown).…”

Cloning, Analysis and Inactivation of the ndhK Gene Encoding a Subunit of NADH Quinone Oxidoreductase from Anabaena PCC 7120

“…The evidence for NDH-1 in cyanobacteria has been obtained from two approaches : biochemical studies and the cloning of genes coding for homologous subunits of complex I. 12 ndh genes have been cloned and sequenced from cyanobacteria (Steinmiiller et al, 1989;Takahashi et al, 1991;Ogawa, a, 1992aEllersiek and Steinmiiller, 1992;Steinmiiller, 1992;Marco et al, 1993;Schluchter et al, 1993). Eleven of these encode homologues of the 14 subunits required for the minimal form of NDH-1.…”