PMID:3087956

Citation	Todd, JA, Roberts, AN, Johnstone, K, Piggot, PJ, Winter, G and Ellar, DJ (1986) Reduced heat resistance of mutant spores after cloning and mutagenesis of the Bacillus subtilis gene encoding penicillin-binding protein 5. J. Bacteriol. 167:257-64
Abstract	Part of the gene encoding penicillin-binding protein 5 from Bacillus subtilis 168 was cloned in Escherichia coli with a synthetic oligonucleotide as a hybridization probe. The gene was designated dacA by analogy with E. coli. The nucleotide sequence was determined, and the predicted molecular mass was 45,594 daltons (412 amino acids). A comparison of the predicted amino acid sequence with that of the E. coli penicillin-binding protein 5 indicated that these enzymes showed about 25% identity. The B. subtilis dacA gene was mutated by integration of a plasmid into the structural gene by homologous recombination. A comparison of the mutant and control strains revealed that (i) the mutant lacked detectable penicillin-binding protein 5, (ii) the D-alanine carboxypeptidase activity of membranes isolated from the mutant was only 5% of that measured in membranes from the control strain, (iii) the mutant cells showed apparently normal morphology only during exponential growth, and after the end of exponential phase the cells became progressively shorter, (iv) the mutant sporulated normally except that the forespore occupied about two-thirds of the mother cell cytoplasm and, during its development, migrated towards the center of the mother cell, and (v) purified mutant spores were 10-fold less heat resistant but possessed normal refractility and morphology. Preliminary chemical analysis indicated that the structure of the cortex of the mutant was different.
Links	PubMed PMC212869
Keywords	Amino Acid Sequence; Bacillus subtilis/cytology; Bacillus subtilis/genetics; Bacillus subtilis/growth & development; Bacillus subtilis/physiology; Bacterial Proteins; Base Sequence; Carboxypeptidases/physiology; Carrier Proteins/genetics; Carrier Proteins/physiology; Cloning, Molecular; Escherichia coli/genetics; Genes; Genes, Bacterial; Hexosyltransferases; Hot Temperature; Muramoylpentapeptide Carboxypeptidase/genetics; Muramoylpentapeptide Carboxypeptidase/physiology; Mutation; Penicillin-Binding Proteins; Peptidyl Transferases; Phenotype; Spores, Bacterial/genetics; Spores, Bacterial/physiology
edit table

Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
BACSU:DACA		GO:0009002: serine-type D-Ala-D-Ala carboxypeptidase activity	ECO:0000247:	UniProtKB:P0AEB2	F		There are several regions of similarity between the amino acid sequences of B. subtilis PBP 5 and E. coli PBP 5 (4) (Fig. 2), accounting for an overall 25% identity. This suggests (8) that these proteins are homologous. The similar regions (Fig. 2) include one around the active-site serine residues (positions 36 and 44 for B. subtilis and E. coli, respectively), extending from positions 7 to 56 in B. subtilis PBP 5 (47% identical amino acids).	complete CACAO 2809
BACSU:DACA	enables	GO:0009002: serine-type D-Ala-D-Ala carboxypeptidase activity	ECO:0000247: sequence alignment evidence used in manual assertion	UniProtKB:P0AEB2	F		Seeded From UniProt	complete

References

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