Evidence that extracellular export of the endoglucanase encoded by egl of Pseudomonas solanacearum occurs by a two-step process involving a lipoprotein intermediate.
Pseudomonas solanacearum is an important phytopathogen that produces a variety of extracellular enzymes. Previous reports suggested that one of these, a 43-kDa beta-1,4-endoglucanase (EGL), is initially synthesized with a 45-residue leader sequence that is removed during export. Experiments with globomycin presented here also suggest that the primary precursor of EGL (ppEGL) has a 45-residue leader sequence but that only the first 19 residues of the leader sequence are removed by signal peptidase II during initial export across the inner membrane. Further analysis suggested that the resultant 46-kDa intermediate precursor (pEGL) is a transient fatty acylated lipoprotein and is located on the periplasmic side of the inner membrane of P. solanacearum. Although Escherichia coli could synthesize ppEGL, modify it with palmitate, and remove the first 19 residues of the leader sequence during export across the inner membrane, only P. solanacearum could export pEGL across the outer membrane and remove the remaining 26 residues of the leader sequence producing the mature, extracellular EGL. The second step of the export process requires export machinery not present in E. coli. To our knowledge this represents the first example of a leader sequence with two distinct parts, one removed during export across the inner membrane and the other removed during export across the outer membrane.
Department of Microbiology, University of Georgia, Athens 30602.
J Z
JZ
Huang
M A
MA
Schell
D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000378:metabolism; D002480:Cellulase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005798:Genes, Bacterial; D005990:Glycerol / Q000378:metabolism; D007700:Kinetics; D008074:Lipoproteins / Q000378:metabolism*; D008954:Models, Biological; D008969:Molecular Sequence Data; D019308:Palmitic Acid; D010169:Palmitic Acids / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals / Q000235:genetics; D011549:Pseudomonas / Q000201:enzymology* / Q000235:genetics
D008074:Lipoproteins; D010169:Palmitic Acids; D021382:Protein Sorting Signals; D019308:Palmitic Acid; D002480:Cellulase; D005990:Glycerol