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Infection and Immunity, April 1999, p. 1844-1852, Vol. 67, No. 4
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Examination of Listeria monocytogenes Intracellular Gene Expression by Using the Green Fluorescent Protein of Aequorea victoria

Nancy E. Freitag,* and Kathleen E. Jacobs

Department of Immunology and Microbiology, Wayne State University School of Medicine, Detroit, Michigan

Received 7 October 1998/Returned for modification 25 November 1998/Accepted 19 January 1999

The ActA protein of Listeria monocytogenes is an essential virulence factor and is required for intracellular bacterial motility and cell-to-cell spread. plcB, cotranscribed with actA, encodes a broad-specificity phospholipase C that contributes to lysis of host cell vacuoles and cell-to-cell spread. Construction of a transcriptional fusion between actA-plcB and the green fluorescent protein gene of Aequorea victoria has facilitated the detailed examination of patterns of actA/plcB expression within infected tissue culture cells. actA/plcB expression began approximately 30 min postinfection and was dependent upon entry of L. monocytogenes into the host cytosol. L. monocytogenes Delta hly mutants, which are unable to escape from host cell vacuoles, did not express actA/plcB at detectable levels within infected tissue culture cells; however, complementation of the hly defect allowed entry of the bacteria into the host cytoplasm and subsequent actA/plcB expression. These results emphasize the ability of L. monocytogenes to sense the different host cell compartment environments encountered during the course of infection and to regulate virulence gene expression in response.

* Corresponding author. Mailing address: Department of Immunology and Microbiology, Wayne State University School of Medicine, Detroit, MI 48201. Phone: (313) 577-1314. Fax: (313) 577-1155. E-mail: nfreitag{at}med.wayne.edu.

Infection and Immunity, April 1999, p. 1844-1852, Vol. 67, No. 4
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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