Open AccessRestriction-modification mediated barriers to exogenous DNA uptake and incorporation employed by Prevotella intermedia
Author(s) -
Christopher D Johnston,
Chelsey A. Skeete,
Alexey Fomenkov,
Richard J. Roberts,
Susan R. Rittling
Publication year - 2017
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0185234
Subject(s) - biology , crispr , prevotella intermedia , intermedia , genetics , computational biology , gene , bacteria , porphyromonas gingivalis , art , performance art , art history
Prevotella intermedia , a major periodontal pathogen, is increasingly implicated in human respiratory tract and cystic fibrosis lung infections. Nevertheless, the specific mechanisms employed by this pathogen remain only partially characterized and poorly understood, largely due to its total lack of genetic accessibility. Here, using S ingle M olecule, R eal- T ime (SMRT) genome and methylome sequencing, bisulfite sequencing, in addition to cloning and restriction analysis, we define the specific genetic barriers to exogenous DNA present in two of the most widespread laboratory strains, P . intermedia ATCC 25611 and P . intermedia Strain 17. We identified and characterized multiple restriction-modification (R-M) systems, some of which are considerably divergent between the two strains. We propose that these R-M systems are the root cause of the P . intermedia transformation barrier. Additionally, we note the presence of conserved Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) systems in both strains, which could provide a further barrier to exogenous DNA uptake and incorporation. This work will provide a valuable resource during the development of a genetic system for P . intermedia , which will be required for fundamental investigation of this organism’s physiology, metabolism, and pathogenesis in human disease.