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Journal of Bacteriology, October 1998, p. 5117-5122, Vol. 180, No. 19
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Formation of Single-Stranded DNA during DNA Transformation of Neisseria gonorrhoeae

Michael S. Chaussee* and Stuart A. Hilldagger

Laboratory of Microbial Structure and Function, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, Montana 59840

Received 15 December 1997/Accepted 3 August 1998

Neisseria gonorrhoeae is naturally competent for DNA transformation. In contrast to other natural prokaryotic DNA transformation systems, single-stranded donor DNA (ssDNA) has not previously been detected during transformation of N. gonorrhoeae. We have reassessed the physical nature of gonococcal transforming DNA by using a sensitive nondenaturing native blotting technique that detects ssDNA. Consistent with previous analyses, we found that the majority of donor DNA remained in the double-stranded form, and only plasmid DNAs that carried the genus-specific DNA uptake sequence were sequestered in a DNase I-resistant state. However, when the DNA was examined under native conditions, S1 nuclease-sensitive ssDNA was identified in all strains tested except for those bacteria that carried the dud-1 mutation. Surprisingly, ssDNA was also found during transformation of N. gonorrhoeae comA mutants, which suggested that ssDNA was initially formed within the periplasm.

* Corresponding author. Mailing address: 903 S. 4th St., Hamilton, MT 59840. Phone: (406) 363-9306. Fax: (406) 363-9204. E-mail: mchaussee{at}nih.gov.

dagger Present address: Department of Biological Sciences, Northern Illinois University, Dekalb, IL 60115.

Journal of Bacteriology, October 1998, p. 5117-5122, Vol. 180, No. 19
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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