Enzymatic activities and functional interdependencies of Bacillus subtilis lipoteichoic acid synthesis enzymes.
Lipoteichoic acid (LTA) is an important cell wall polymer in Gram-positive bacteria. The enzyme responsible for polyglycerolphosphate LTA synthesis is LtaS, first described in Staphylococcus aureus. Four LtaS orthologues, LtaS(BS) , YfnI, YqgS and YvgJ, are present in Bacillus subtilis. Using an in vitro enzyme assay, we determined that all four proteins are Mn(2+) -dependent metal enzymes that use phosphatidylglycerol as a substrate. We show that LtaS(BS) , YfnI and YqgS can produce polymers, suggesting that these three proteins are bona-fide LTA synthases while YvgJ functions as an LTA primase, as indicated by the accumulation of a GroP-Glc(2) -DAG glycolipid. Western blot analysis of LTA produced by ltaS(BS) , yfnI, yqgS and yvgJ single, triple and the quadruple mutant, showed that LTA production was only abolished in the quadruple and the YvgJ-only expressing mutant. B. subtilis strains expressing YfnI in the absence of LtaS(BS) produced LTA of retarded mobility, presumably caused by an increase in chain length as suggested by a structural analysis of purified LTA. Taken together, the presented results indicate that the mere presence or absence of LTA cannot account for cell division and sporulation defects observed in the absence of individual enzymes and revealed an unexpected enzymatic interdependency of LtaS-type proteins in B. subtilis.
Section of Microbiology Division of Molecular Biosciences, Imperial College London, South Kensington Campus, London SW7 2AZ, UK.
Mirka E
ME
Wörmann
Rebecca M
RM
Corrigan
Peter J
PJ
Simpson
Steve J
SJ
Matthews
Angelika
A
Gründling
D001412:Bacillus subtilis / Q000187:drug effects / Q000201:enzymology*; D001426:Bacterial Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002855:Chromatography, Thin Layer; D004789:Enzyme Activation / Q000187:drug effects; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism; D005816:Genetic Complementation Test; D006017:Glycolipids / Q000737:chemistry / Q000302:isolation & purification; D006868:Hydrolysis / Q000187:drug effects; D007700:Kinetics; D008070:Lipopolysaccharides / Q000096:biosynthesis* / Q000302:isolation & purification; D009682:Magnetic Resonance Spectroscopy; D008345:Manganese / Q000494:pharmacology; D008954:Models, Biological; D008970:Molecular Weight; D009154:Mutation / Q000235:genetics; D010715:Phosphatidylglycerols / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013211:Staphylococcus aureus / Q000187:drug effects / Q000378:metabolism; D013379:Substrate Specificity / Q000187:drug effects; D013682:Teichoic Acids / Q000096:biosynthesis* / Q000302:isolation & purification
D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D006017:Glycolipids; D008070:Lipopolysaccharides; D010715:Phosphatidylglycerols; D011994:Recombinant Proteins; D013682:Teichoic Acids; D008345:Manganese; C009900:lipoteichoic acid
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