Natural Transformation of Helicobacter pylori Involves the Integration of Short DNA Fragments Interrupted by Gaps of Variable Size

Open Access
Publisher Website
Google Scholar
Abstract
Helicobacter pylori are gram-negative bacteria notable for their high level of genetic diversity and plasticity, features that may play a key role in the organism's ability to colonize the human stomach. Homeologous natural transformation, a key contributor to genomic diversification, has been well-described for H. pylori. To examine the mechanisms involved, we performed restriction analysis and sequencing of recombination products to characterize the length, fragmentation, and position of DNA imported via natural transformation. Our analysis revealed DNA imports of small size (1,300 bp, 95% confidence limits 950–1850 bp) with instances of substantial asymmetry in relation to selectable antibiotic-resistance markers. We also observed clustering of imported DNA endpoints, suggesting a possible role for restriction endonucleases in limiting recombination length. Additionally, we observed gaps in integrated DNA and found evidence suggesting that these gaps are the result of two or more separate strand invasions. Taken together, these observations support a system of highly efficient short-fragment recombination involving multiple recombination events within a single locus. Helicobacter pylori are gram-negative bacteria that have been implicated in human diseases after decades of persistence in the stomach. Known for its high level of genetic diversity, H. pylori is competent to undergo natural transformation, a process in which donor DNA is integrated into the recipient chromosome. To examine the mechanisms involved, we analyzed the DNA imported via natural transformation in an experimental model system. We found variation in the average length of imported DNA fragments, with asymmetry with respect to a selectable marker. We also found evidence that strain-specific restriction endonucleases may limit recombination length. Additionally, we observed gaps in the integrated DNA and provide evidence that these gaps are the result of separate strand invasions. Together, our observations support a highly efficient system of short-fragment recombination involving multiple recombination events within a small region of the chromosome. This helps explain how bacteria are able to employ genetic recombination to efficiently generate and maintain genomic diversification within the population—a feature that helps H. pylori persistently colonize the harsh environment of the human stomach.