Lipid modification of prelipoproteins is dispensable for growth but essential for efficient protein secretion in Bacillus subtilis: characterization of the Lgt gene.
We have identified and characterized the Igt gene of Bacillus subtilis. The prelipoprotein diacylglycerol transferase enzyme (Lgt) catalyses the first reaction in lipomodification of bacterial lipoproteins. Inactivation of Igt in B. subtilis by a nonsense mutation (prs-11 mutation) or by disruption was shown here to abolish lipomodification of prelipoproteins completely, as well as the cleavage of signal peptide. However, unlike in Gram-negative bacteria, the Igt mutants of B. subtilis were fully viable. In agreement with this observation, studies of two lipoproteins, PrsA and BlaP, indicated that non-lipomodified precursors of these proteins were functional and translocated across the cytoplasmic membrane. However, there was release of both precursors from cells, resulting in a reduced level of the cell-bound form. We have shown that the reduced level of the PrsA lipoprotein, a foldase involved in protein secretion, caused impaired protein secretion, a prominent phenotype of Igt mutants. There was no indication that non-lipomodified PrsA displayed reduced activity.
Leskelä
S
S
Laboratory of Vaccine Development, National Public Health Institute, Helsinki, Finland.
Wahlström
E
E
Kontinen
V P
VP
Sarvas
M
M
D001412:Bacillus subtilis / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D001426:Bacterial Proteins; D002874:Chromosome Mapping; D010450:Endopeptidases / Q000378:metabolism; D005798:Genes, Bacterial; D008074:Lipoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D011498:Protein Precursors / Q000378:metabolism; D013381:Subtilisins / Q000378:metabolism; D014166:Transferases / Q000235:genetics*; D000516:alpha-Amylases / Q000378:metabolism
D001426:Bacterial Proteins; D008074:Lipoproteins; D008565:Membrane Proteins; D011498:Protein Precursors; C082160:prsA protein, bacteria; D014166:Transferases; C042308:phosphatidylglycerol glyceryl transferase; D000516:alpha-Amylases; D010450:Endopeptidases; D013381:Subtilisins