title: Analysis of spontaneous suppressor mutants from the photomixotrophically grown pmgA-disrupted mutant in the cyanobacterium Synechocystis sp. PCC 6803.
abstract: The pmgA-disrupted (ΔpmgA) mutant in the cyanobacterium Synechocystis sp. PCC 6803 suffers severe growth inhibition under photomixotrophic conditions. In order to elucidate the key factors enabling the cells to grow under photomixotrophic conditions, we isolated spontaneous suppressor mutants from the ΔpmgA mutant derived from a single colony. When the ΔpmgA mutant was spread on a BG11 agar plate supplemented with glucose, colonies of suppressor mutants appeared after the bleaching of the background cells. We identified the mutation site of these suppressor mutants and found that 11 mutants out of 13 had a mutation in genes related to the type 1 NAD(P)H dehydrogenase (NDH-1) complex. Among them, eight mutants had mutations within the ndhF3 (sll1732) gene: R32stop, W62stop, V147I, G266V, G354W, G586C, and deletion of 7 bp within the coding region. One mutant had one base insertion in the putative -10 box of the ndhC (slr1279) gene, leading to the decrease in the transcripts of the ndhCKJ operon. Two mutants had one base insertion and deletion in the coding region of cupA (sll1734), which is co-transcribed with ndhF3 and ndhD3 and comprises together a form of NDH-1 complex (NDH-1MS complex) involved in inducible high-affinity CO2 uptake. The results indicate that the loss of the activity of this complex effectively rescues the ΔpmgA mutant under photomixotrophic condition with 1 % CO2. However, little difference among WT and mutants was observed in the activities ascribed to the NDH-1MS complex, i.e., CO2 uptake and cyclic electron transport. This may suggest that the NDH-1MS complex has the third, currently unknown function under photomixotrophic conditions.
authors: Graduate School of Science and Engineering, Saitama University, Saitama, 338-8570, Japan.
authors: Yoshiki
authors: Y
authors: Nishijima
authors: Nodai Genome Research Center, Tokyo University of Agriculture, Tokyo, 156-8502, Japan.
authors: Yu
authors: Y
authors: Kanesaki
authors: Nodai Genome Research Center, Tokyo University of Agriculture, Tokyo, 156-8502, Japan.|Department of Bioscience, Tokyo University of Agriculture, Tokyo, 156-8502, Japan.|CREST, Japan Science and Technology Agency (JST), Saitama, 332-0012, Japan.
authors: Hirofumi
authors: H
authors: Yoshikawa
authors: Faculty of Education and Integrated Arts and Sciences, Waseda University, Tokyo, 162-8480, Japan.|Japan Society for the Promotion of Science, Tokyo, 102-0083, Japan.
authors: Takako
authors: T
authors: Ogawa
authors: Faculty of Education and Integrated Arts and Sciences, Waseda University, Tokyo, 162-8480, Japan. sonoike@waseda.jp.
authors: Kintake
authors: K
authors: Sonoike
authors: Graduate School of Science and Engineering, Saitama University, Saitama, 338-8570, Japan.
authors: Yoshitaka
authors: Y
authors: Nishiyama
authors: Graduate School of Science and Engineering, Saitama University, Saitama, 338-8570, Japan.|CREST, Japan Science and Technology Agency (JST), Saitama, 332-0012, Japan.
authors: Yukako
authors: Y
authors: Hihara
mesh_terms: D004579:Electron Transport; D008027:Light; D009154:Mutation; D010084:Oxidation-Reduction; D010788:Photosynthesis; D046939:Synechocystis / Q000235:genetics* / Q000502:physiology / Q000528:radiation effects
keywords: Cyanobacteria; Glucose tolerance; NAD(P)H dehydrogenase; Photomixotrophic growth; Suppressor mutation; ndhF3