Project description:In this study, disruption and overexpression of sigD were performed in Corynebacterium glutamicum and analyzed by transcriptome sequencing (RNA-seq) to understand the SigD regulon in C. glutamicum. For the effect of sigD overexpression, the relative abundance of mRNA was compared in WT(pVWEx1-sigD) without IPTG or with 50 M of IPTG. For the effect of sigD disruption, the abundance was compared between the sigD disrupted mutant and the wild type strain.
Project description:(Coryno)mycolate is a α-branched, β-hydroxylated long-chain fatty acid specifically synthesized in bacteria in the suborder Corynebacterineae of the phylum Actinobacteria. It forms an outer membrane and functions as a permeability barrier conferring pathogenic mycobacteria to resistance to antibiotics. Whereas mycolate biosynthetic pathway has been intensively studied, the studies on the transcriptional regulation of genes involved in the pathway are limited. Here, we report that the previously uncharacterized extracytoplasmic function σ factor, σD, is a key regulator of the mycolate synthetic genes in Corynebacterium glutamicum in the suborder. Chromatin immunoprecipitation with microarray analysis detected σD-binding regions in the genome, establishing a consensus promoter sequence for σD recognition. The σD regulon comprises acyl-CoA carboxylase subunits, acyl-AMP ligase, polyketide synthase, and mycolyltransferases, all of which are involved in mycolate synthesis. Actually, deletion or overexpression of sigD encoding σD modified the amount of extractable mycolate. Immediately downstream of sigD, rsdA encoded anti-σD and was under the control of a σD-dependent promoter. Another σD regulon member, L,D-transpeptidase, conferred lysozyme resistance. Thus, σD modifies cell wall composition and enhances mycolate synthesis to provide resistance to environmental stress.
Project description:To identify genes which are differentially expressed in Corynebacterium glutamicum in the cg2699 deletion strain, we performed DNA microarray analyses of C. glutamicum Δcg2699 compared to the WT.
Project description:To identify genes which are differentially expressed in Corynebacterium glutamicum in the cg2460 deletion strain, we performed DNA microarray analyses of C. glutamicum Δcg2460 compared to the WT.
Project description:Corynebacterium glutamicum GlxR is a homolog of the cAMP receptor protein. Although over 200 GlxR binding sites in the C. glutamicum genome are predicted in silico, studies on the GlxR physiological function have been hindered by the severe growth defects of a glxR mutant. This study comprehensively 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. Moreover, ChIP-chip analyses showed that GlxR was still able to interact with its target sites in a deletion mutant of cyaB, the sole adenylate cyclase gene in the genome, even though binding affinity was markedly decreased.
Project description:To identify genes which are differentially expressed in Corynebacterium glutamicum chassis C1 in comparison to the prophage free strain MB001, we performed DNA microarray analyses of C. glutamicum C1 against MB001. For this purpose RNA was isolated from cells cultivated in CGXII minimal medium with 2% glucose (w v-1) and harvested in the exponential growth phase at an OD600 of 5. Four biological replicates were performed.