GONUTS has been updated to MW1.31 Most things seem to be working but be sure to report problems.

Have any questions? Please email us at ecoliwiki@gmail.com

PMID:16352845

From GONUTS
Jump to: navigation, search
Citation

De Lay, NR and Cronan, JE (2006) Gene-specific random mutagenesis of Escherichia coli in vivo: isolation of temperature-sensitive mutations in the acyl carrier protein of fatty acid synthesis. J. Bacteriol. 188:287-96

Abstract

Acyl carrier proteins (ACPs) are very small acidic proteins that play a key role in fatty acid and complex lipid synthesis. Moreover, recent data indicate that the acyl carrier protein of Escherichia coli has a large protein interaction network that extends beyond lipid synthesis. Despite extensive efforts over many years, no temperature-sensitive mutants with mutations in the structural gene (acpP) that encodes ACP have been isolated. We report the isolation of three such mutants by a new approach that utilizes error-prone PCR mutagenesis, overlap extension PCR, and phage lambda Red-mediated homologous recombination and that should be generally applicable. These mutants plus other experiments demonstrate that ACP function is essential for the growth of E. coli. Each of the mutants was efficiently modified with the phosphopantetheinyl moiety essential for the function of ACP in lipid synthesis, and thus lack of function at the nonpermissive temperature cannot be attributed to a lack of prosthetic group attachment. All of the mutant proteins were largely stable at the nonpermissive temperature except the A68T/N73D mutant protein. Fatty acid synthesis in strains that carried the D38V or A68T/N73D mutations was inhibited upon a shift to the nonpermissive temperature and in the latter case declined to a small percentage of the rate of the wild-type strain.

Links

PubMed PMC1317600 Online version:10.1128/JB.188.1.287-296.2006

Keywords

Acyl Carrier Protein/genetics; Acyl Carrier Protein/metabolism; Amino Acid Substitution; Apoproteins/genetics; Apoproteins/metabolism; Escherichia coli K12/enzymology; Escherichia coli K12/genetics; Escherichia coli K12/growth & development; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; Fatty Acid Synthetase Complex, Type II; Fatty Acids/biosynthesis; Genes, Essential; Genetic Techniques; Models, Molecular; Mutagenesis; Polymerase Chain Reaction; Recombination, Genetic; Temperature

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

ECOLI:ACP

acts_upstream_of_or_within

GO:0006633: fatty acid biosynthetic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:ACP

acts_upstream_of_or_within

GO:0009245: lipid A biosynthetic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:ACP

acts_upstream_of_or_within

GO:0008610: lipid biosynthetic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:ACP

enables

GO:0031177: phosphopantetheine binding

ECO:0000315: mutant phenotype evidence used in manual assertion

F

Seeded From UniProt

complete

ECOLI:ACP

involved_in

GO:0009245: lipid A biosynthetic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:ACP

involved_in

GO:0008610: lipid biosynthetic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

ECOLI:ACP

involved_in

GO:0006633: fatty acid biosynthetic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

See Help:References for how to manage references in GONUTS.