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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
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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