title: Coordinated synthesis of the two ClpB isoforms improves the ability of Escherichia coli to survive thermal stress.
abstract: Eubacteria synthesize a full-length (ClpB95) and a N-terminally truncated (ClpB80) version of the ClpB disaggregase owing to the presence of a translation initiation site within the clpB transcript. Why these two isoforms have been evolutionary conserved is poorly understood. Here, we constructed a series of E. coli strains and plasmids allowing production of the ClpB95/ClpB80 pair, ClpB95 alone, or ClpB80 alone from near physiological concentrations to a 6-10-fold excess over normal cellular levels. We found that although overexpressed ClpB95 or ClpB80 can independently restore basal thermotolerance to DeltaclpB cells, strains expressing ClpB80 from the clpB chromosomal locus do not exhibit increased resistance to thermal killing at 50 degrees C relative to clpB null cells. Furthermore, synthesis of physiological levels of ClpB95 is less effective than coordinated expression of ClpB95/ClpB80 in protecting E. coli from thermal killing. These results provide an explanation for the conservation of the two ClpB isoforms in eubacteria and are consistent with the fact that wild type E. coli maintains the ClpB80 to ClpB95 ratio at a nearly constant value of 0.4-0.5 under a variety of stress conditions.
authors.affiliation: Department of Chemical Engineering, University of Washington, Box 351750, Seattle, WA 98195, USA.
authors.forename: I-Ting
authors.initials: IT
authors.lastname: Chow
authors.forename: François
authors.initials: F
authors.lastname: Baneyx
mesh_terms: D017124:Conserved Sequence; D049071:Endopeptidase Clp; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000235:genetics; D019143:Evolution, Molecular; D017353:Gene Deletion; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics; D006358:Hot Temperature; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics
chemical_list: D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins; D020033:Protein Isoforms; D049071:Endopeptidase Clp; C069775:ClpB protein, E coli