THE use of lacZ gene fusions, producing a hybrid protein containing an amino terminus specified by a target gene fused to the functional carboxy terminus of β-galactosidase, has facilitated the study of protein targeting in various organisms. One of the best characterized fusions in Eschcrichia coli is Φ(lamB-lacZ)42-l(Hyb), which produces a hybrid protein with the signal sequence and 181 N-terminal amino acids of the exported protein LamB, attached to LacZ1. In common with other LacZ hybrids (reviewed in ref. 2), the LamB-LacZ(42-l) protein is poorly exported from E. coli, conferring a Lac+ phenotype. β-Galactosidase activity decreases markedly when cells producing the LamB-LacZ protein are grown at 42°C or when a heat-shock response is induced at lower temperatures by overproducing heat-shock factor RpoH3, indicating the LacZ hybrids are being efficiently targeted to the cell envelope. We now report that the heat-shock proteins DnaK and GroEL can, in sufficient amounts, decrease β-galactosidase activity and facilitate the export of lacZ-hybrid proteins.
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