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PMID:18067544

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Citation

Nanamiya, H, Kasai, K, Nozawa, A, Yun, CS, Narisawa, T, Murakami, K, Natori, Y, Kawamura, F and Tozawa, Y (2008) Identification and functional analysis of novel (p)ppGpp synthetase genes in Bacillus subtilis. Mol. Microbiol. 67:291-304

Abstract

Bacterial alarmone (p)ppGpp, is a global regulator responsible for the stringent control. Two homologous (p)ppGpp synthetases, RelA and SpoT, have been identified and characterized in Escherichia coli, whereas Gram-positive bacteria such as Bacillus subtilis have been thought to possess only a single RelA-SpoT enzyme. We have now identified two genes, yjbM and ywaC, in B. subtilis that encode a novel type of alarmone synthetase. The predicted products of these genes are relatively small proteins ( approximately 25 kDa) that correspond to the (p)ppGpp synthetase domain of RelA-SpoT family members. A database survey revealed that genes homologous to yjbM and ywaC are conserved in certain bacteria belonging to Firmicutes or Actinobacteria phyla but not in other phyla such as Proteobacteria. We designated the proteins as small alarmone synthetases (SASs) to distinguish them from RelA-SpoT proteins. The (p)ppGpp synthetase function of YjbM and YwaC was confirmed by genetic complementation analysis and by in vitro assay of enzyme activity. Molecular genetic analysis also revealed that ywaC is induced by alkaline shock, resulting in the transient accumulation of ppGpp. The SAS proteins thus likely function in the biosynthesis of alarmone with a mode of action distinct from that of RelA-SpoT homologues.

Links

PubMed Online version:10.1111/j.1365-2958.2007.06018.x

Keywords

Amino Acid Sequence; Arginine/analogs & derivatives; Arginine/metabolism; Bacillus subtilis/classification; Bacillus subtilis/enzymology; Bacillus subtilis/genetics; Bacterial Proteins/chemistry; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Escherichia coli/genetics; Escherichia coli Proteins/chemistry; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; Gene Expression Regulation, Bacterial; Genetic Complementation Test; Guanosine Tetraphosphate/biosynthesis; Ligases/chemistry; Ligases/genetics; Ligases/metabolism; Ligases/physiology; Molecular Sequence Data; Mutation; Phylogeny; Sequence Alignment; Transcription, Genetic

Significance

Annotations

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

BACSU:YJBM

GO:0008728: GTP diphosphokinase activity

ECO:0000314:

F

Figure 3C: Shows in vitro YjbM activity with use of radiolabeled ATP. Figure shows pppGpp synthesis in presence of YjbM.

complete

BACSU:YWAC

GO:0008728: GTP diphosphokinase activity

ECO:0000314:

F

Figure 3C: Shows pppGpp synthesis from GTP and radiolabeled ATP in presence of purified YwaC.

complete


See also

References

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