Isolation and characterization of an Escherichia coli mutant having temperature-sensitive farnesyl diphosphate synthase.
The screening of a collection of highly mutagenized strains of Escherichia coli for defects in isoprenoid synthesis led to the isolation of a mutant that had temperature-sensitive farnesyl diphosphate synthase. The defective gene, named ispA, was mapped at about min 10 on the E. coli chromosome, and the gene order was shown to be tsx-ispA-lon. The mutant ispA gene was transferred to the E. coli strain with a defined genetic background by P1 transduction for investigation of its function. The in vitro activity of farnesyl diphosphate synthase of the mutant was 21% of that of the wild-type strain at 30 degrees C and 5% of that at 40 degrees C. At 42 degrees C the ubiquinone level was lower (66% of normal) in the mutant than in the wild-type strain, whereas at 30 degrees C the level in the mutant was almost equal to that in the wild-type strain. The polyprenyl phosphate level was slightly higher in the mutant than in the wild-type strain at 30 degrees C and almost the same in both strains at 42 degrees C. The mutant had no obvious phenotype regarding its growth properties.
Fujisaki
S
S
Department of Chemistry, Faculty of General Education, Gifu University, Japan.
Nishino
T
T
Katsuki
H
H
Hara
H
H
Nishimura
Y
Y
Hirota
Y
Y
D019883:Alkyl and Aryl Transferases; D002851:Chromatography, High Pressure Liquid; D002874:Chromosome Mapping; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D051228:Geranyltranstransferase; D045782:Hemiterpenes; D009154:Mutation; D009943:Organophosphorus Compounds / Q000378:metabolism; D011106:Polyisoprenyl Phosphates / Q000378:metabolism*; D012717:Sesquiterpenes; D013696:Temperature; D014166:Transferases / Q000235:genetics* / Q000378:metabolism; D014451:Ubiquinone / Q000378:metabolism
D045782:Hemiterpenes; D009943:Organophosphorus Compounds; D011106:Polyisoprenyl Phosphates; D012717:Sesquiterpenes; D014451:Ubiquinone; C004809:isopentenyl pyrophosphate; C004808:farnesyl pyrophosphate; D014166:Transferases; D019883:Alkyl and Aryl Transferases; D051228:Geranyltranstransferase