title: Role of Corynebacterium glutamicum sprA encoding a serine protease in glxR-mediated global gene regulation.
abstract: The global regulator glxR of Corynebacterium glutamicum is involved in many cellular activities. Considering its role, the GlxR protein likely interacts with other proteins to obtain, maintain, and control its activity. To isolate proteins interacting with GlxR, we used a two-hybrid system with GlxR as the bait. Subsequently, the partner, a subtilisin-like serine protease, was isolated from a C. glutamicum genomic library. Unlike glxR, which showed constitutive expression, the expression of sprA, encoding a serine protease, was maximal in the log phase. Purified His6-SprA protein underwent self-proteolysis and proteolyzed purified GlxR. The proteolytic action of SprA on GlxR was not observed in the presence of cyclic adenosine monophosphate, which modulates GlxR activity. The C. glutamicum sprA deletion mutant (ΔsprA) and sprA-overexpressing (P180-sprA) strains showed reduced growth. The activity of isocitrate dehydrogenase (a tricarboxylic acid cycle enzyme) in these strains decreased to 30-50% of that in the wild-type strain. In the P180-sprA strain, proteins involved in diverse cellular functions such as energy and carbon metabolism (NCgl2809), nitrogen metabolism (NCgl0049), methylation reactions (NCgl0719), and peptidoglycan biosynthesis (NCgl1267), as well as stress, starvation, and survival (NCgl0938) were affected and showed decreased transcription. Taken together, these data suggest that SprA, as a serine protease, performs a novel regulatory role not only in glxR-mediated gene expression but also in other areas of cell physiology. In addition, the tight control of SprA and GlxR availability may indicate their importance in global gene regulation.
authors.affiliation: Department of Biotechnology and Bioinformatics, Korea University, Sejong-ro, Sejong-si, Korea.
authors.forename: Eun-Ji
authors.initials: EJ
authors.lastname: Hong
authors.affiliation: Department of Biotechnology and Bioinformatics, Korea University, Sejong-ro, Sejong-si, Korea.
authors.forename: Joon-Song
authors.initials: JS
authors.lastname: Park
authors.affiliation: Department of Oriental Medicine, Semyung University, Checheon, Chungbuk, Korea.
authors.forename: Younhee
authors.initials: Y
authors.lastname: Kim
authors.affiliation: Department of Biotechnology and Bioinformatics, Korea University, Sejong-ro, Sejong-si, Korea.
authors.forename: Heung-Shick
authors.initials: HS
authors.lastname: Lee
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D048230:Corynebacterium glutamicum / Q000201:enzymology / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D005947:Glucose / Q000235:genetics / Q000378:metabolism; D059748:Proteolysis; D012097:Repressor Proteins / Q000235:genetics*; D057057:Serine Proteases / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D001426:Bacterial Proteins; D012097:Repressor Proteins; D057057:Serine Proteases; D005947:Glucose