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Infection and Immunity, November 2008, p. 5110-5119, Vol. 76, No. 11
0019-9567/08/$08.00+0     doi:10.1128/IAI.00445-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Bacterial Conjugation in the Cytoplasm of Mouse Cells{triangledown}

Yin Mei Lim,1,{dagger} Ad J. C. de Groof,2,{dagger},{ddagger} Mrinal K. Bhattacharjee,3 David H. Figurski,4 and Eric A. Schon1,2*

Departments of Genetics and Development,1 Neurology,2 Microbiology, Columbia University, New York, New York 10032,4 Department of Chemistry and Biochemistry, Long Island University, Brooklyn, New York 112013

Received 10 April 2008/ Returned for modification 19 May 2008/ Accepted 21 August 2008

Intracellular pathogenic organisms such as salmonellae and shigellae are able to evade the effects of many antibiotics because the drugs are not able to penetrate the plasma membrane. In addition, these bacteria may be able to transfer genes within cells while protected from the action of drugs. The primary mode by which virulence and antibiotic resistance genes are spread is bacterial conjugation. Salmonellae have been shown to be competent for conjugation in the vacuoles of cultured mammalian cells. We now show that the conjugation machinery is also functional in the mammalian cytosol. Specially constructed Escherichia coli strains expressing Shigella flexneri plasmid and chromosomal virulence factors for escape from vacuoles and synthesizing the invasin protein from Yersinia pseudotuberculosis to enhance cellular entry were able to enter 3T3 cells and escape from the phagocytic vacuole. One bacterial strain (the donor) of each pair to be introduced sequentially into mammalian cells had a conjugative plasmid. We found that this plasmid could be transferred at high frequency. Conjugation in the cytoplasm of cells may well be a general phenomenon.

* Corresponding author. Mailing address: Columbia University Medical Center, 1150 St. Nicholas Ave., Room 303A, New York, NY 10032. Phone: (212) 851-5532. Fax: (212) 305-3986. E-mail: eas3{at}columbia.edu

{triangledown} Published ahead of print on 2 September 2008.

Editor: J. B. Bliska

{dagger} Y.M.L. and A.J.C.D.G. contributed equally to this work.

{ddagger} Present address: Department of Cell Biology, Nijmegen Centre for Molecular Life Sciences, Radboud University Medical Centre, Geert Grooteplein 28, 6525 GA Nijmegen, The Netherlands.

Infection and Immunity, November 2008, p. 5110-5119, Vol. 76, No. 11
0019-9567/08/$08.00+0     doi:10.1128/IAI.00445-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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