TY - JOUR
T1 - Generation and analysis of an ICE R391 deletion library identifies genes involved in the element encoded UV-inducible cell-sensitising function
AU - Armshaw, Patricia
AU - Pembroke, J. Tony
N1 - © 2013 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.
PY - 2013/5
Y1 - 2013/5
N2 - ICE R391, a prototype member of the SXT/R391 family of site-specific integrative conjugative elements (ICEs), frequently isolated from enterobacterial pathogens, exhibits an unusual, recA-dependent, UV-inducible, cell-sensitising function. This significantly decreases postirradiation cell survival rates in Escherichia coli host cells, a trait that would at first appear to be counterproductive in terms of adaptation to stress conditions. Construction and screening of a complete ICE R391 deletion library in E. coli identified three ICE R391 genes, orfs90/91, encoding a putative transcriptional enhancer, and orf43, encoding a putative type IV secretion system outer membrane-associated conjugative transfer protein, in the cell-sensitising function. Cloning and complementation of these genes confirmed their involvement in UV sensitising. Expression of both orfs90/91 and orf43 in wild-type E. coli indicated that orf43 encodes a cytotoxic gene product upon up-regulation. Deletion of the orf43 homologue in SXT, s050, also abolished its associated UV sensitisation. We hypothesise that ICE R391 and other members of the SXT/R391 family display decreased survival rates upon exposure to UV irradiation through the induction of orf43.
AB - ICE R391, a prototype member of the SXT/R391 family of site-specific integrative conjugative elements (ICEs), frequently isolated from enterobacterial pathogens, exhibits an unusual, recA-dependent, UV-inducible, cell-sensitising function. This significantly decreases postirradiation cell survival rates in Escherichia coli host cells, a trait that would at first appear to be counterproductive in terms of adaptation to stress conditions. Construction and screening of a complete ICE R391 deletion library in E. coli identified three ICE R391 genes, orfs90/91, encoding a putative transcriptional enhancer, and orf43, encoding a putative type IV secretion system outer membrane-associated conjugative transfer protein, in the cell-sensitising function. Cloning and complementation of these genes confirmed their involvement in UV sensitising. Expression of both orfs90/91 and orf43 in wild-type E. coli indicated that orf43 encodes a cytotoxic gene product upon up-regulation. Deletion of the orf43 homologue in SXT, s050, also abolished its associated UV sensitisation. We hypothesise that ICE R391 and other members of the SXT/R391 family display decreased survival rates upon exposure to UV irradiation through the induction of orf43.
KW - Conjugative transfer
KW - SXT/R391-like elements
KW - Type IV secretion systems
KW - UV-sensitising
UR - http://www.scopus.com/inward/record.url?scp=84876333032&partnerID=8YFLogxK
U2 - 10.1111/1574-6968.12107
DO - 10.1111/1574-6968.12107
M3 - Article
C2 - 23413978
AN - SCOPUS:84876333032
SN - 0378-1097
VL - 342
SP - 45
EP - 53
JO - FEMS Microbiology Letters
JF - FEMS Microbiology Letters
IS - 1
ER -