Enhanced export of beta-galactosidase fusion proteins in prlF mutants is Lon dependent.
We have used fusions of the outer membrane protein LamB to beta-galactosidase (encoded by lacZ) to study the protein export process. This LamB-LacZ hybrid protein blocks export when synthesized at high levels, as evidenced by inducer (maltose) sensitivity, a phenomenon termed LacZ hybrid jamming. The prlF1 mutation relieves LacZ hybrid jamming and allows localization of the fusion protein to a noncytoplasmic compartment. prlF1 and similar alleles are gain-of-function mutations. Null mutations in this gene confer no obvious phenotypes. Extragenic suppressors of a gain-of-function prlF allele have been isolated in order to understand how this gene product affects the export process. The suppressors are all lon null mutations, and they are epistatic to all prlF phenotypes tested. Lon protease activity has been measured in prlF1 cells and shown to be increased. However, the synthesis of Lon is not increased in a prlF1 background, suggesting a previously unidentified mechanism of Lon activation. Further analysis reveals that prlF1 activates degradation of cytoplasmically localized precursors in a Lon protease-dependent manner. It is proposed that accumulation of precursors during conditions of hybrid protein jamming titrates an essential export component(s), possibly a chaperone. Increased Lon-dependent precursor degradation would free this component, thus allowing increased protein export under jamming conditions.
Snyder
W B
WB
Department of Molecular Biology, Princeton University, New Jersey 08544.
Silhavy
T J
TJ
D049069:ATP-Dependent Proteases; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins / Q000378:metabolism*; D001692:Biological Transport; D002451:Cell Compartmentation; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins; D016153:Genes, Suppressor; D006360:Heat-Shock Proteins; D008242:Lysogeny; D018272:Porins; D049070:Protease La; D011991:Receptors, Virus / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000378:metabolism*; D001616:beta-Galactosidase / Q000235:genetics
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins; D018272:Porins; D011991:Receptors, Virus; D011993:Recombinant Fusion Proteins; C016532:maltoporins; D001616:beta-Galactosidase; D049069:ATP-Dependent Proteases; D012697:Serine Endopeptidases; C483567:Lon protein, E coli; D049070:Protease La