Heat shock response in Corynebacterium glutamicum was characterized by whole-genome expression analysis using a DNA microarray. It was indicated that heat shock response of C. glutamicum included not only upregulation of heat shock protein (HSP) genes encoding molecular chaperones and ATP-dependent proteases, but it also increased and decreased expression of more than 300 genes related to disparate physiological functions. An extracytoplasmic-function sigma factor, SigH, was upregulated by heat shock. The SigH regulon was defined by gene expression profiling using sigH-disrupted and overexpressing strains in conjunction with mapping of transcription initiation sites. A total of 45 genes, including HSP genes and genes involved in oxidative stress response, were identified as the SigH regulon. Expression of some HSP genes was also upregulated by deletion of the transcriptional regulators HspR and HrcA. HspR represses expression of the clpB and dnaK operons, and HrcA represses expression of groESL1 and groEL2. SigH was shown to play an important role in regulation of heat shock response in concert with HspR and HrcA, but its role is likely restricted to only a part of the regulation of C. glutamicum heat shock response. Upregulation of 18 genes encoding transcriptional regulators by heat shock suggests a complex regulatory network of heat shock response in C. glutamicum.

Corynebacterium glutamicum GlxR is a cyclic AMP (cAMP) receptor protein-type regulator. Although over 200 GlxR-binding sites in the C. glutamicum genome are predicted in silico, studies on the physiological function of GlxR have been hindered by the severe growth defects of a glxR mutant. This study identified the GlxR regulon by chromatin immunoprecipitation in conjunction with microarray (ChIP-chip) analyses. In total, 209 regions were detected as in vivo GlxR-binding sites. In vitro binding assays and promoter-reporter assays demonstrated that GlxR directly activates expression of genes for aerobic respiration, ATP synthesis, and glycolysis and that it is required for expression of genes for cell separation and mechanosensitive channels. GlxR also directly represses a citrate uptake gene in the presence of citrate. Moreover, ChIP-chip analyses showed that GlxR was still able to interact with its target sites in a mutant with a deletion of cyaB, the sole adenylate cyclase gene in the genome, even though binding affinity was markedly decreased. Thus, GlxR is physiologically functional at the relatively low cAMP levels in the cyaB mutant, allowing the cyaB mutant to grow much better than the glxR mutant.

The sigB gene of Corynebacterium glutamicum encodes a group 2 sigma factor of RNA polymerase. Under conditions of oxygen deprivation, the sigB gene is upregulated and cells exhibit high productivity of organic acids as a result of an elevated glucose consumption rate. Using DNA microarray and quantitative reverse transcription-PCR (RT-PCR) analyses, we found that sigB disruption led to reduced transcript levels of genes involved in the metabolism of glucose into organic acids. This in turn resulted in retardation of glucose consumption by cells under conditions of oxygen deprivation. These results indicate that SigB is involved in positive regulation of glucose metabolism genes and enhances glucose consumption under conditions of oxygen deprivation. Moreover, sigB disruption reduced the transcript levels of genes involved in various cellular functions, including the glucose metabolism genes not only in the growth-arrested cells under conditions of oxygen deprivation but also in the cells during aerobic exponential growth, suggesting that SigB functions as another vegetative sigma factor.

Excessive light conditions repressed the levels of mRNAs accumulation of multiple Lhc genes encoding light-harvesting chlorophyll-a/b (LHC) proteins of photosystem (PS)II in the unicellular green alga, Chlamydomonas reinhardtii. The light intensity required for the repression tended to decrease with lowering temperature or CO 2 concentration. The responses of six LhcII genes encoding the major LHC (LHCII) proteins and two genes (Lhcb4 and Lhcb5) encoding the minor LHC proteins of PSII (CP29 and CP26) were similar. The results indicate that the expression of these Lhc genes is coordinately repressed when the energy input through the antenna systems exceeds the requirement for CO 2 assimilation. The Lhc mRNA level repressed under high-light conditions was partially recovered by adding the electron transport inhibitor 3-(3,4-dichlorophenyl)-1,1-dimethylurea, suggesting that redox signaling via photosynthetic electron carriers is involved in the gene regulation. However, the mRNA level was still considerably lower under high-light than under low-light conditions even in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea. Repression of the Lhc genes by high light was prominent even in the mutants deficient in the reaction center(s) of PSII or both PSI and PSII. The results indicate that two alternative processes are involved in the repression of Lhc genes under high-light conditions, one of which is independent of the photosynthetic reaction centers and electron transport events.Photosynthesis is regulated at various levels in response to fluctuating light intensity under various ambient temperature and nutrient conditions. The proper responses to the various environmental cues are necessary for photosynthetic plants to use light energy efficiently and to protect themselves from photoinhibitory damage caused by excessive irradiance (Aro et al., 1993;Long et al., 1994;Osmond, 1994). Excessive light energy absorbed by chlorophyll is dissipated by non-radiative processes (Crofts and Yerkes, 1994;Horton et al., 1996;Gilmore, 1997) and is properly distributed between two photosystems (PS) by state transition (Allen, 1995;Gal et al., 1997), whereas the energy input is regulated by changes in the size of the light-harvesting antenna systems to modulate the optical cross section.Light-harvesting chlorophyll a/b (LHC)II proteins, which are major components of light-harvesting antennae of PSII in higher plants and green algae, typically change their abundance in response to the intensity of irradiance (Anderson et al., 1988(Anderson et al., , 1995. Under stress and intense light, enhanced amounts of reactive oxygen species will react with proteins and lipids, not only in chloroplasts but also in the cytosol, and will induce various types of photodamage. Therefore, the quality and quantity control of the LHC protein complex is required to avoid photodamage by alleviating excitation energy pressure. Although the LHC protein complex could be controlled by various mechanisms including pigment synthesis, the repression of the...

The Lhcb gene family in green plants encodes several light-harvesting Chl a/b-binding (LHC) proteins that collect and transfer light energy to the reaction centers of PSII. We comprehensively characterized the Lhcb gene family in the unicellular green alga, Chlamydomonas reinhardtii, using the expressed sequence tag (EST) databases. A total of 699 among over 15,000 ESTs related to the Lhcb genes were assigned to eight, including four new, genes that we isolated and sequenced here. A sequence comparison revealed that six of the Lhcb genes from C. reinhardtii correspond to the major LHC (LHCII) proteins from higher plants, and that the other two genes (Lhcb4 and Lhcb5) correspond to the minor LHC proteins (CP29 and CP26). No ESTs corresponding to another minor LHC protein (CP24) were found. The six LHCII proteins in C. reinhardtii cannot be assigned to any of the three types proposed for higher plants (Lhcb1-Lhcb3), but were classified as follows: Type I is encoded by LhcII-1.1, LhcII-1.2 and LhcII-1.3, and Types II, III and IV are encoded by LhcII-2, LhcII-3 and LhcII-4, respectively. These findings suggest that the ancestral LHC protein diverged into LHCII, CP29 and CP26 before, and that LHCII diverged into multiple types after the phylogenetic separation of green algae and higher plants.