Glutamate and cyclic AMP regulate the expression of galactokinase in Mycobacterium smegmatis.
It was found that Mycobacterium smegmatis is unable to utilize galactose as the sole carbon source because the sugar alone cannot induce galactokinase. However, galactokinase was induced by glutamate alone, and was further stimulated by galactose. Rifampicin completely inhibited the glutamate-mediated expression of galK in both the absence and presence of galactose. Extracellular cAMP stimulated the expression of the enzyme only in the presence of glutamate plus galactose. The galK gene from M. smegmatis, including its upstream promoter region, was cloned in a plasmid in Escherichia coli. The expression of kinase from these clones in E. coli was dependent on cAMP and its receptor protein (CRP). The expression of UDP-galactose 4-epimerase was constitutive. This and other evidence suggests that the galK gene is not linked to galT and galE in the mycobacterial genome. In a glutamate-independent galactose-utilizing mutant (gin-1 mutant) of M. smegmatis, galK was expressed in the absence of both galactose and glutamate, while in the presence of galactose this expression was increased twofold in the absence of glutamate and fourfold in its presence. Extracellularly added cAMP reduced the expression of the enzyme in the presence of galactose plus glutamate nearly to the basal level. It is proposed that in M. smegmatis the galK gene is expressed from two different promoters; the expression from one promoter is dependent on glutamate but not on galactose and cAMP, while that from the other requires all three components. The role of galactose is possibly to derepress the latter promoter.
Department of Biochemistry, Bose InstituteAcharya J.C. Bose Birth Centenary Building, P-1/12, CIT Scheme VII M, Calcutta 700054India.
Santanu
S
Raychaudhuri
Department of Biochemistry, Bose InstituteAcharya J.C. Bose Birth Centenary Building, P-1/12, CIT Scheme VII M, Calcutta 700054India.
Madhumita
M
Basu
Department of Biochemistry, Bose InstituteAcharya J.C. Bose Birth Centenary Building, P-1/12, CIT Scheme VII M, Calcutta 700054India.
Nitai C
NC
Mandal
D001693:Biological Transport, Active; D000242:Cyclic AMP / Q000494:pharmacology / Q000502:physiology*; D002373:Cyclic AMP Receptor Protein / Q000502:physiology; D004357:Drug Synergism; D005686:Galactokinase / Q000096:biosynthesis* / Q000235:genetics; D005690:Galactose / Q000378:metabolism / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D018698:Glutamic Acid / Q000502:physiology*; D009161:Mycobacterium / Q000201:enzymology* / Q000235:genetics; D014158:Transcription, Genetic; D014534:UDPglucose 4-Epimerase / Q000096:biosynthesis
D002373:Cyclic AMP Receptor Protein; D018698:Glutamic Acid; D000242:Cyclic AMP; D005686:Galactokinase; D014534:UDPglucose 4-Epimerase; D005690:Galactose