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Journal of Bacteriology, September 2000, p. 5225-5230, Vol. 182, No. 18
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Characterization of Escherichia coli DNA Lesions Generated within J774 Macrophages

Eliana Schlosser-Silverman,1 Maya Elgrably-Weiss,1 Ilan Rosenshine,1 Ron Kohen,2 and Shoshy Altuvia1,*

Department of Molecular Genetics and Biotechnology, The Hebrew University-Hadassah Medical School,1 and Department of Pharmaceutics, School of Pharmacy,2 The Hebrew University, 91120 Jerusalem, Israel

Received 23 February 2000/Accepted 1 June 2000

Macrophages are armed with multiple oxygen-dependent and -independent bactericidal properties. However, the respiratory burst, generating reactive oxygen species, is believed to be a major cause of bacterial killing. We exploited the susceptibility of Escherichia coli in macrophages to characterize the effects of the respiratory burst on intracellular bacteria. We show that E. coli strains recovered from J774 macrophages exhibit high rates of mutations. We report that the DNA damage generated inside macrophages includes DNA strand breaks and the modification 8-oxo-2'-deoxyguanosine, which are typical oxidative lesions. Interestingly, we found that under these conditions, early in the infection the majority of E. coli cells are viable but gene expression is inhibited. Our findings demonstrate that macrophages can cause severe DNA damage to intracellular bacteria. Our results also suggest that protection against the macrophage-induced DNA damage is an important component of the bacterial defense mechanism within macrophages.

* Corresponding author. Mailing address: Department of Molecular Genetics and Biotechnology, The Hebrew University-Hadassah Medical School, 91120 Jerusalem, Israel. Phone: 972-2-675-7212. Fax: 972-2-678-4010. E-mail: shoshy{at}cc.huji.ac.il.

Journal of Bacteriology, September 2000, p. 5225-5230, Vol. 182, No. 18
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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