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Microbiology 154 (2008), 2680-2688; DOI  10.1099/mic.0.2008/019075-0IMMEDIATE OPEN ACCESS ARTICLE
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Microbiology 154 (2008), 2680-2688; DOI  10.1099/mic.0.2008/019075-0
© 2008 Society for General Microbiology

The Salmonella SPI-2 effector SseJ exhibits eukaryotic activator-dependent phospholipase A and glycerophospholipid : cholesterol acyltransferase activity

Nadine S. Lossi1, Nathalie Rolhion1, Anthony I. Magee2, Cliona Boyle1 and David W. Holden1

1 Centre for Molecular Microbiology and Infection, Imperial College London, Armstrong Road, London SW7 2AZ, UK
2 National Heart and Lung Institute, Imperial College London, London SW7 2AZ, UK

Correspondence
David W. Holden
d.holden{at}imperial.ac.uk

Intracellular replication of Salmonella enterica serovar Typhimurium within membrane-bound compartments, called Salmonella-containing vacuoles, depends on the activities of several effector proteins translocated by the Salmonella pathogenicity island 2 (SPI-2)-encoded type III secretion system. The SPI-2 effector protein SseJ shows similarity at the amino acid level to several GDSL lipases with glycerophospholipid : cholesterol acyltransferase (GCAT) activity. In this study, we show that catalytic serine-dependent phospholipase A (PLA) and GCAT activity of recombinant SseJ is potentiated by factor(s) present in HeLa cells, RAW macrophages and Saccharomyces cerevisiae. SseJ activity was enhanced with increasing amounts of, or preincubation with, eukaryotic cell extracts. Analysis of the activating factor(s) shows that it is soluble and heat- and protease-sensitive. We conclude that PLA and GCAT activities of SseJ are potentiated by proteinaceous eukaryotic factor(s).


Abbreviations: CE, cholesterol ester; DPPC, 1,2-dipalmitoylphosphatidylcholine; FFA, free fatty acids; GCAT, glycerophospholipid : , cholesterol acyltransferase; GST, glutathione-S-transferase; 1-MPLPC, 1-monopalmitoyllysophosphatidylcholine; PLA, phospholipase A; PLA2, phospholipase A2; PNPB, para-nitrophenyl butyrate; PNS, postnuclear supernatant; SCV, Salmonella-containing vacuole; SPI-2, Salmonella pathogenicity island 2; T3SS, type III secretion system







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