Enterococcus faecalis multi-drug resistance transporters: application for antibiotic discovery.
Using bioinformatics approaches, 34 potential multidrug resistance (MDR) transporter sequences representing 4 different transporter families were identified in the unannotated Enterococcus faecalis database (TIGR). A functional genomics campaign generating single-gene insertional disruptions revealed several genes whose absence confers significant hypersensitivities to known antimicrobials. We constructed specific strains, disrupted in a variety of previously unpublished, putative MDR transporter genes, as tools to improve the success of whole-cell antimicrobial screening and discovery. Each of the potential transporters was inactivated at the gene level and then phenotypically characterized, both with single disruption mutants and with 2-gene mutants built upon a delta norA deleted strain background.
Davis
D R
DR
Phytera, Inc, Worcester, MA 01605, USA.
McAlpine
J B
JB
Pazoles
C J
CJ
Talbot
M K
MK
Alder
E A
EA
White
C
C
Jonas
B M
BM
Murray
B E
BE
Weinstock
G M
GM
Rogers
B L
BL
D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D019992:Databases as Topic; D015195:Drug Design; D004352:Drug Resistance, Microbial; D013293:Enterococcus faecalis / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D017353:Gene Deletion; D023281:Genomics; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D016296:Mutagenesis; D013329:Structure-Activity Relationship
D018528:ATP-Binding Cassette Transporters; D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D002352:Carrier Proteins; D027425:ATP-Binding Cassette, Sub-Family C Proteins; C075040:NorA protein, Staphylococcus