title: fusB is an allele of nadD, encoding nicotinate mononucleotide adenylyltransferase in Escherichia coli.
abstract: Isolation of the temperature-sensitive Escherichia coli mutant 72c has been described previously. The mutant allele was named fusB and causes a pleiotropic phenotype, the most striking features of which, besides temperature sensitivity, are the inability to grow on synthetic medium and supersensitivity to trimethoprim, an antibiotic that inhibits the C1 metabolism. This work shows that the fusB mutation is a frameshift mutation in the nadD gene that encodes nicotinate mononucleotide adenylyltransferase. The frameshift leads to a change of the last 10 amino acids and an addition of 17 amino acids. This lesion, renamed nadD72, leads to very little NAD+ and NADPH synthesis at the permissive temperature and essentially no synthesis at the non-permissive temperature. As a comparison, a new mutation in the nadD gene, with an amino acid change in the ATP-binding site, has been isolated. Its NAD+ synthesis is decreased at 30 degrees C but the level is still sufficient to support normal growth. At 42 degrees C, NAD+ synthesis is reduced further, which leads to temperature sensitivity on minimal medium. This mutation was designated nadD74. Thus, a small decrease in NAD+ levels affects ability to grow on minimal medium at 42 degrees C, while a large decrease leads to a more pleiotropic phenotype.
authors.affiliation: Department of Microbiology, Stockholm University, S-106 91 Stockholm, Sweden.
authors.forename: Martin
authors.initials: M
authors.lastname: Stancek
authors.affiliation: Department of Microbiology, Stockholm University, S-106 91 Stockholm, Sweden.
authors.forename: Leif A
authors.initials: LA
authors.lastname: Isaksson
authors.affiliation: Department of Microbiology, Stockholm University, S-106 91 Stockholm, Sweden.
authors.forename: Monica
authors.initials: M
authors.lastname: Rydén-Aulin
mesh_terms: D000483:Alleles; D002855:Chromatography, Thin Layer; D003470:Culture Media; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D015723:Gene Library; D009154:Mutation; D009612:Nicotinamide-Nucleotide Adenylyltransferase; D009713:Nucleotidyltransferases / Q000096:biosynthesis* / Q000235:genetics; D010641:Phenotype; D013696:Temperature
chemical_list: D003470:Culture Media; D009713:Nucleotidyltransferases; D009612:Nicotinamide-Nucleotide Adenylyltransferase; C038480:nicotinic acid mononucleotide adenylyltransferase