The reason(s) for glucose sensitivity in certain cyanobacterial strains is poorly understood. Inactivation of genes encoding the putative sensor kinase Hik31 in Synechocystis sp. strain PCC 6803 resulted in a mutant unable to grow in the presence of d-glucose. Sensitivities to d-glucose, its analogue 2-deoxy-d-glucose, and fructose, were alleviated in mutants in which glcP, encoding the glucose transporter, was inactivated. These data indicate that permeation of these substrates is required to inflict cell death. The mutant Δhik31, and the glucose-sensitive strain of Synechocystis, do not possess glucokinase activity, although a transcript originating from glk, encoding glucokinase, is present. Inactivation of glk led to severe sensitivity to glucose, indicating that the presence of glucose itself, within the cells, inflicted this sensitivity. On the other hand, sensitivity to 2-deoxy-d-glucose was lower in Δglk, thus distinguishing between the effect of glucose itself and that of its analogue, which, in the absence of glucokinase activity, may not be phosphorylated. Addition of glucose led to a small rise in glucose-6-phosphate dehydrogenase activity in the wild type, but constitutive activity was observed in the Δhik31 mutant regardless of the presence of glucose. Microarray analyses showed only small changes in the abundance of global transcripts in Synechocystis following glucose addition, but the transcription levels of several genes, including icfG, but not glk, were strongly affected by inactivation of hik31. The mechanism(s) whereby Hik31 is involved in glucose sensing and response is discussed.
BeufL, BeduS, DurandM. C, JosetF.
1994; A protein involved in co-ordinated regulation of inorganic carbon and glucose metabolism in the facultative photoautrophic cyanobacterium Synechocystis PCC 6803. Plant Mol Biol 25:855–864[CrossRef]
BloyeS. A, SilmanN. J, MannN. H, CarrN. G.
1992; Bicarbonate concentration by Synechocystis PCC 6803 modulation of protein phosphorylation and inorganic carbon transport by glucose. Plant Physiol 99:601–606[CrossRef]
FloresE, SchmettererG.
1986; Interaction of fructose with the glucose permease of the cyanobacterium Synechocystis sp. strain PCC 6803. J Bacteriol 166:693–696
GleasonF. K.
1996; Glucose-6-phosphate dehydrogenase from the cyanobacterium, Anabaena sp. PCC 7120: purification and kinetics of redox modulation. Arch Biochem Biophys 334:277–283[CrossRef]
GranotD, DalN.
1997; Sugar induced cell death in yeast is dependent on the rate of sugar phosphorylation as determined by Arabidopsis thaliana hexokinase. Cell Death Differ 4:555–559[CrossRef]
HagenK. D, MeeksJ. C.
2001; The unique cyanobacterial protein OpcA is an allosteric effector of glucose-6-phosphate dehydrogenase in Nostoc punctiforme ATCC 29133. J Biol Chem 276:11477–11486[CrossRef]
HiharaY, IkeuchiM.
1997; Mutation in a novel gene required for photomixotrophic growth leads to enhanced photoautotrophic growth of Synechocystis sp. PCC 6803. Photosynth Res 53:243–252[CrossRef]
HiharaY, SonoikeK, IkeuchiM.
1998; A novel gene, pmgA , specifically regulates photosystem stoichiometry in the cyanobacterium Synechocystis sp. PCC 6803 in response to high light. Plant Physiol 117:1206–1216
HiharaY, MuramatsuM, NakamuraK, SonoikeK.
2004; A cyanobacterial gene encoding an ortholog of Pirin is induced under stress conditions. FEBS Lett 574:101–105[CrossRef]
HsiaoH. Y, HeQ. F,
van WaasbergenL. G, GrossmanA. R.
2004; Control of photosynthetic and high-light-responsive genes by the histidine kinase DspA: negative and positive regulation and interactions between signal transduction pathways. J Bacteriol 186:3882–3888[CrossRef]
IwasakiW, MiyatakeH, EbiharaA, MikiK.
2004; Crystallization and preliminary X-ray crystallographic studies of the small form of glucose-inhibited division protein A from Thermus thermophilus HB8. Acta Cryst D60:515–517
KaplanA, BadgerM. R, BerryJ. A.
1980; Photosynthesis and intracellular inorganic carbon pool in the blue-green algae Anabaena variabilis : response to external CO[sub]2[/sub] concentration. Planta 149:219–226[CrossRef]
KatohH, HaginoN, GrossmanA. R, OgawaT.
2001; Genes essential to iron transport in the cyanobacterium Synechocystis sp. strain PCC 6803. J Bacteriol 183:2779–2784[CrossRef]
KnowlesV. L, PlaxtonW. C.
2003; From genome to enzyme: analysis of key glycolytic and oxidative pentose-phosphate pathway enzymes in the cyanobacterium Synechocystis sp. PCC 6803. Plant Cell Physiol 44:758–763[CrossRef]
KoksharovaO, SchubertM, ShestakovS, CerffR.
1998; Genetic and biochemical evidence for distinct key functions of two highly divergent GAPDH genes in catabolic and anabolic carbon flow of the cyanobacterium Synechocystis sp. PCC 6803. Plant Mol Biol 36:183–194[CrossRef]
KulmaA, VilladsenD, CampbellD. G, MeekS. E. M. E, HarthillJ, NielsenT. H, MacKintoshC.
2004; Phosphorylation and 14-3-3 binding of Arabidopsis 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase. Plant J 37:654–667[CrossRef]
LemaireK, Van de VeldeS, Van DijckP, TheveleinJ. M.
2004; Glucose and sucrose act as agonist and mannose as antagonist ligands of the G protein-coupled receptor Gpr1 in the yeast Saccharomyces cerevisiae . Mol Cell 16:293–299[CrossRef]
MinH. T, GoldenS. S.
2000; A new circadian class 2 gene, opcA , whose product is important for reductant production at night in Synechococcus elongatus PCC 7942. J Bacteriol 182:6214–6221[CrossRef]
MooreB. L. Z, RollandF, HallQ, ChengW. H, LiuY. X, HwangI, JonesT, SheenJ.
2003; Role of the Arabidopsis glucose sensor HXK1 in nutrient, light, and hormonal signaling. Science 300:332–336[CrossRef]
MoriyaH, JohnstonM.
2004; Glucose sensing and signaling in Saccharomyces cerevisiae through the Rgt2 glucose sensor and casein kinase I. Proc Natl Acad Sci U S A 101:1572–1577[CrossRef]
RinconA. M, CodonA. C, CastrejonF, BenitezT.
2001; Improved properties of baker's yeast mutants resistant to 2-deoxy-d-glucose. Appl Environ Microbiol 67:4279–4285[CrossRef]
RippkaR, DeruellesJ, WaterburyJ.-B, HerdmanM, StanierR.-Y.
1979; Genetic assignments, strain histories and properties of pure cultures of cyanobacteria. J Gen Microbiol 111:1–61[CrossRef]
RomanowskiM. J, BonannoJ. B, BurleyS. K.
2002; Crystal structure of the Escherichia coli glucose-inhibited division protein B (GidB) reveals a methyltransferase fold. Proteins 47:563–567[CrossRef]
RyuJ. Y, SongJ. Y, LeeJ. M, JeongS. W, ChowW. S, ChoiS. B, PogsonB. J, ParkY. I.
2004; Glucose-induced expression of carotenoid biosynthesis genes in the dark is mediated by cytosolic pH in the cyanobacterium Synechocystis sp PCC 6803. J Biol Chem 279:25320–25325[CrossRef]
ShiL, BischoffK. M, KennellyP. J.
1999; The icfG gene cluster of Synechocystis sp. strain PCC 6803 encodes an Rsb/Spo-like protein kinase, protein phosphatase, and two phosphoproteins. J Bacteriol 181:4761–4767
SinghA. K, ShermanL. A.
2005; Pleiotropic effect of a histidine kinase on carbohydrate metabolism in Synechocystis sp. strain PCC 6803 and its requirement for heterotrophic growth. J Bacteriol 187:2368–2376[CrossRef]
SundaramS, KarakayaH, ScanlanD. J, MannN. H.
1998; Multiple oligomeric forms of glucose-6-phosphate dehydrogenase in cyanobacteria and the role of OpcA in the assembly process. Microbiology 144:1549–1556[CrossRef]
TuC.-J, ShragerJ, BurnapR. L, PostierB. L, GrossmanA. R.
2004; Consequences of a deletion in dspA on transcript accumulation in Synechocystis sp. strain PCC 6803. J Bacteriol 186:3889–3902[CrossRef]
von MeyenburgK, JorgensenB. B, NielsenJ, HansenF. G.
1982; Promoters of the atp operon coding for the membrane-bound ATP synthase of Escherichia coli mapped by Tn 10 insertion mutations. Mol Gen Genet 188:240–248[CrossRef]
YangC, HuaQ, ShimizuK.
2002; Integration of the information from gene expression and metabolic fluxes for the analysis of the regulatory mechanisms in Synechocystis . Appl Microbiol Biotechnol 58:813–822[CrossRef]
ZhangC. C, JeanjeanR, JosetF.
1998; Obligate phototrophy in cyanobacteria: more than a lack of sugar transport. FEMS Microbiol Lett 161:285–292[CrossRef]