title: Kinetics of biosynthesis of iron-regulated membrane proteins in Escherichia coli.
abstract: Using biological iron chelators to control specifically iron availability to Escherichia coli K-12 in conjunction with radioactive pulse-labels, we examined the biosynthesis of six iron-regulated membrane proteins. Iron deprivation induced the synthesis of five proteins, which had molecular weights of 83,000 (83K), 81K (Fep), 78K (TonA), 74K (Cir), and 25K. The kinetics of induction were the same in entA and entA(+) strains, but were affected by the initial iron availability in the media. Iron-poor cells induced rapidly (half-time, 10 min), whereas iron-rich cells began induction after a lag and showed a slower induction half-time (30 min). Within this general pattern of induction after iron deprivation, several different kinetic patterns were apparent. The 83K, 81K, and 74K proteins were coordinately controlled under all of the conditions examined. The 78K and 25K proteins were regulated differently. The synthesis of a previously unrecognized 90K inner membrane protein was inhibited by iron deprivation and stimulated by iron repletion. Both ferrichrome and ferric enterobactin completely repressed 81K and 74K synthesis when the siderophores were supplied at concentrations of 5 muM in vivo (half-time, 2.5 min). At concentrations less than 5 muM, however, both siderophores repressed synthesis only temporarily; the duration of repression was proportional to the amount of ferric siderophore added. The half-lives of the 81K and 74K mRNAs, as measured by rifampin treatment, were 1.2 and 1.6 min, respectively. The results of this study suggest that enteric bacteria are capable of instantaneously detecting and reacting to fluctuations in the extracellular iron concentration and that they store iron during periods of iron repletion for utilization during periods of iron stress. Neither iron storage nor iron regulation of envelope protein synthesis is dependent on the ability of the bacteria to form heme.
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis*; D004758:Enterobactin / Q000096:biosynthesis / Q000494:pharmacology; D004926:Escherichia coli / Q000378:metabolism*; D005291:Ferrichrome / Q000494:pharmacology; D006207:Half-Life; D007501:Iron / Q000494:pharmacology*; D007700:Kinetics; D008565:Membrane Proteins / Q000096:biosynthesis*; D008970:Molecular Weight; D012329:RNA, Bacterial / Q000378:metabolism; D012333:RNA, Messenger / Q000494:pharmacology
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; D005291:Ferrichrome; D004758:Enterobactin; D007501:Iron