Combined, functional genomic-biochemical approach to intermediary metabolism: interaction of acivicin, a glutamine amidotransferase inhibitor, with Escherichia coli K-12.
Acivicin, a modified amino acid natural product, is a glutamine analog. Thus, it might interfere with metabolism by hindering glutamine transport, formation, or usage in processes such as transamidation and translation. This molecule prevented the growth of Escherichia coli in minimal medium unless the medium was supplemented with a purine or histidine, suggesting that the HisHF enzyme, a glutamine amidotransferase, was the target of acivicin action. This enzyme, purified from E. coli, was inhibited by low concentrations of acivicin. Acivicin inhibition was overcome by the presence of three distinct genetic regions when harbored on multicopy plasmids. Comprehensive transcript profiling using DNA microarrays indicated that histidine biosynthesis was the predominant process blocked by acivicin. The response to acivicin, however, was quite complex, suggesting that acivicin inhibition resonated through more than a single cellular process.
Smulski
D R
DR
Biochemical Science and Engineering, Central Research and Development, DuPont Company, Wilmington, DE 19880-0173, USA.
Huang
L L
LL
McCluskey
M P
MP
Reeve
M J
MJ
Vollmer
A C
AC
Van Dyk
T K
TK
LaRossa
R A
RA
D000619:Aminohydrolases / Q000235:genetics / Q000378:metabolism; D000878:Anthranilate Synthase; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001667:Binding, Competitive; D003470:Culture Media; D004791:Enzyme Inhibitors / Q000378:metabolism / Q000494:pharmacology*; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000379:methods; D005973:Glutamine / Q000378:metabolism; D007093:Imidazoles / Q000378:metabolism; D007555:Isoxazoles / Q000378:metabolism / Q000494:pharmacology*; D019884:Nitrogenous Group Transferases / Q000037:antagonists & inhibitors* / Q000235:genetics* / Q000378:metabolism; D012265:Ribonucleotides / Q000378:metabolism; D000637:Transaminases / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D003470:Culture Media; D004791:Enzyme Inhibitors; D007093:Imidazoles; D007555:Isoxazoles; D012265:Ribonucleotides; D005973:Glutamine; C081348:N(1)-((5'-phosphoribulosyl)formimino)-5-aminoimidazo-4-carboxamide ribonucleotide; D019884:Nitrogenous Group Transferases; D000637:Transaminases; C053430:histidinol-phosphate aminotransferase; C087227:imidazole glycerol phosphate synthase; D000619:Aminohydrolases; D000878:Anthranilate Synthase; C020655:anthranilate synthase, glutamine amidotransferase subunit; C006303:acivicin