title: Involvement of cyclic AMP receptor protein in regulation of the rmf gene encoding the ribosome modulation factor in Escherichia coli.
abstract: The decrease in overall translation in stationary-phase Escherichia coli is accompanied with the formation of functionally inactive 100S ribosomes mediated by the ribosome modulation factor (RMF). At present, however, little is known regarding the regulation of stationary-phase-coupled RMF expression. In the course of a systematic screening of regulation targets of DNA-binding transcription factors from E. coli, we realized that CRP (cyclic AMP [cAMP] receptor protein), the global regulator for carbon source utilization, participates in regulation of some ribosomal protein genes, including the rmf gene. In this study, we carried out detailed analysis of the regulation of the RMF gene by cAMP-CRP. The cAMP-dependent binding of CRP to the rmf gene promoter was confirmed by gel shift and DNase I footprinting assays. By using a reporter assay system, the expression level of RMF was found to decrease in the crp knockout mutant, indicating the involvement of CRP as an activator of the rmf promoter. In good agreement with the reduction of rmf promoter activity, we observed decreases in RMF production and 100S ribosome dimerization in the absence of CRP. Taken together, we propose that CRP regulates transcription activation of the rmf gene for formation of 100S ribosome dimers. Physiological roles of CRP involvement in RMF production are discussed.
authors.affiliation: Department of Frontier Bioscience, Hosei University, Koganei, Tokyo.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Shimada
authors.forename: Hideji
authors.initials: H
authors.lastname: Yoshida
authors.forename: Akira
authors.initials: A
authors.lastname: Ishihama
mesh_terms: D002373:Cyclic AMP Receptor Protein / Q000235:genetics / Q000378:metabolism*; D018983:DNA Footprinting; D003850:Deoxyribonuclease I; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D024202:Electrophoretic Mobility Shift Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics / Q000502:physiology; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D002373:Cyclic AMP Receptor Protein; D029968:Escherichia coli Proteins; D012269:Ribosomal Proteins; C063545:ribosome modulation factor, E coli; D003850:Deoxyribonuclease I