title: Isolation and mutation site determination of the temperature-sensitive murB mutants of Staphylococcus aureus.
abstract: The murB gene encodes UDP-N-acetylenolpyruvylglucosamine reductase and functions in bacterial peptidoglycan biosynthesis. A plasmid carrying the murB gene restored the temperature-sensitive growth of six Staphylococcus aureus mutants, in which peptidoglycan biosynthesis stopped at a restrictive temperature. Specific activity of UDP-N-acetylenolpyruvylglucosamine reductase in extracts from the mutants was lower than that from wild-type cells. Nucleotide sequence determination revealed that each mutant had a single amino acid substitution in the murB gene and five of six mutations were located within domain 3, where the proposed substrate binding site is located. These results suggest that the murB gene is essential for growth of S. aureus and that domain 3 is important for the MurB activity.
authors.affiliation: Graduate School of Pharmaceutical Sciences, The University of Tokyo, 3-1, 7-Chome, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Miki
authors.initials: M
authors.lastname: Matsuo
authors.forename: Kenji
authors.initials: K
authors.lastname: Kurokawa
authors.forename: Satoshi
authors.initials: S
authors.lastname: Nishida
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Haruto
authors.initials: H
authors.lastname: Takimura
authors.forename: Chikara
authors.initials: C
authors.lastname: Kaito
authors.forename: Norio
authors.initials: N
authors.lastname: Fukuhara
authors.forename: Hideki
authors.initials: H
authors.lastname: Maki
authors.forename: Kenji
authors.initials: K
authors.lastname: Miura
authors.forename: Kazuhisa
authors.initials: K
authors.lastname: Murakami
authors.forename: Kazuhisa
authors.initials: K
authors.lastname: Sekimizu
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites / Q000235:genetics; D002237:Carbohydrate Dehydrogenases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D002473:Cell Wall / Q000201:enzymology / Q000648:ultrastructure; D005816:Genetic Complementation Test; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009154:Mutation; D010457:Peptidoglycan / Q000096:biosynthesis; D010641:Phenotype; D017434:Protein Structure, Tertiary; D013211:Staphylococcus aureus / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D010457:Peptidoglycan; D002237:Carbohydrate Dehydrogenases; C023215:UDP-N-acetylmuramate dehydrogenase