1887

Abstract

, the agent of Legionnaires’ disease, is an intracellular parasite of aquatic protozoans and human macrophages. The type II protein secretion system of the Gram-negative organism promotes intracellular infection. A lipase activity and a -nitrophenylphosphorylcholine (pNPPC) hydrolytic activity are two of the factors that are diminished in type II secretion mutants. The lipase activity was found to include free fatty acid release from di- and triacylglycerol substrates, in addition to the previously reported cleavage of monoacylglycerol. In a number of other bacterial systems, the release of -nitrophenol from pNPPC is due to a phospholipase C. In an attempt to identify exoproteins that potentiate intracellular infection, three genes were identified and mutated in strain 130b that were predicted to encode either a secreted lipase or a phospholipase C. The first two genes, which were designated and , encoded proteins containing the lipase consensus sequence [LIV]-X-[LIVFY]-[LIVMST]-G-[HYWV]-S-X-G-[GSTAC]. Mutations in in particular reduced supernatant activity against mono- and triacylglycerols. However, loss of and/or did not impair the ability of to infect amoebae or U937 cell macrophages. The third gene, which was denoted , encoded a protein that was highly homologous with a phospholipase C from . Inactivation of diminished secreted pNPPC hydrolase activity but did not influence infection of host cells. Taken together, these data indicate that has multiple lipases and possibly several phospholipase C enzymes but that LipA, LipB and PlcA are not among those exoproteins required for optimal intracellular infection.

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2002-07-01
2019-10-18
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