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Impaired synthesis and secretion of SopA in Salmonella Typhimurium dam mutants

Giacomodonato, Mónica N. and Sarnacki, Sebastián H. and Noto Llana, Mariángeles and García Cattaneo, Alejandra S. and Uzzau, Sergio and Rubino, Salvatore and Cerquetti, María Cristina (2009) Impaired synthesis and secretion of SopA in Salmonella Typhimurium dam mutants. FEMS Microbiology Letters, Vol. 292 (1), p. 71-77. eISSN 1574-6968. Article.

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DOI: 10.1111/j.1574-6968.2008.01473.x

Abstract

DNA adenine methylation regulates virulence gene expression in certain bacteria, including Salmonella Typhimurium. The aim of this study was to investigate the involvement of DNA adenine methylase (Dam) methylation in the expression and secretion of the SPI-1 effector protein SopA. For this purpose, SopA–FLAG-tagged wild-type and dam strains of Salmonella Typhimurium were constructed. The expression and secretion of SopA were determined in bacterial culture and in intracellular bacteria recovered from infected HEp-2 epithelial cells. Bacterial culture supernatants and pellets were used to investigate secreted proteins and cell-associated proteins, respectively. Western blot and quantitative reverse transcriptase PCR analysis showed that the dam mutant expresses lower levels of SopA than the wild-type strain. Interestingly, the strain lacking Dam synthesizes SopA under nonpermissive conditions (28 °C). In addition, SopA secretion was drastically impaired in the dam mutant. In vivo experiments showed that the intracellular Salmonella dam mutant synthesizes SopA although in lower amounts than the wild-type strain. Taken together, our results suggest that Dam methylation modulates the expression and secretion of SopA in Salmonella Typhimurium.

Item Type:Article
ID Code:1503
Status:Published
Refereed:Yes
Uncontrolled Keywords:Salmonella Typhimurium, SopA, DNA adenine methylase (Dam)
Subjects:Area 06 - Scienze mediche > MED/07 Microbiologia e microbiologia clinica
Divisions:001 Università di Sassari > 01 Dipartimenti > Scienze biomediche
Publisher:Blackwell / Wiley
eISSN:1574-6968
Copyright Holders:© 2009 Federation of European Microbiological Societies
Deposited On:18 Aug 2009 10:05

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