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PMID:18463693

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Citation

Hett, EC, Chao, MC, Deng, LL and Rubin, EJ (2008) A mycobacterial enzyme essential for cell division synergizes with resuscitation-promoting factor. PLoS Pathog. 4:e1000001

Abstract

The final stage of bacterial cell division requires the activity of one or more enzymes capable of degrading the layers of peptidoglycan connecting two recently developed daughter cells. Although this is a key step in cell division and is required by all peptidoglycan-containing bacteria, little is known about how these potentially lethal enzymes are regulated. It is likely that regulation is mediated, at least partly, through protein-protein interactions. Two lytic transglycosylases of mycobacteria, known as resuscitation-promoting factor B and E (RpfB and RpfE), have previously been shown to interact with the peptidoglycan-hydrolyzing endopeptidase, Rpf-interacting protein A (RipA). These proteins may form a complex at the septum of dividing bacteria. To investigate the function of this potential complex, we generated depletion strains in M. smegmatis. Here we show that, while depletion of rpfB has no effect on viability or morphology, ripA depletion results in a marked decrease in growth and formation of long, branched chains. These growth and morphological defects could be functionally complemented by the M. tuberculosis ripA orthologue (rv1477), but not by another ripA-like orthologue (rv1478). Depletion of ripA also resulted in increased susceptibility to the cell wall-targeting beta-lactams. Furthermore, we demonstrate that RipA has hydrolytic activity towards several cell wall substrates and synergizes with RpfB. These data reveal the unusual essentiality of a peptidoglycan hydrolase and suggest a novel protein-protein interaction as one way of regulating its activity.

Links

PubMed PMC2262848 Online version:10.1371/journal.ppat.1000001

Keywords

Aconitate Hydratase/physiology; Anti-Bacterial Agents/pharmacology; Bacterial Proteins/physiology; Cell Division/physiology; Cell Wall/enzymology; Cytokines/physiology; Drug Synergism; Gene Expression Regulation, Bacterial; Gene Expression Regulation, Enzymologic; Gene Silencing; Genes, Bacterial; Microbial Sensitivity Tests; Microbial Viability; Mycobacterium smegmatis/drug effects; Mycobacterium smegmatis/enzymology; Mycobacterium smegmatis/genetics; Mycobacterium tuberculosis/enzymology; N-Acetylmuramoyl-L-alanine Amidase/physiology; Recombinant Proteins/biosynthesis

Significance

Annotations

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

MYCS2:RIPA

involved_in

GO:0016998: cell wall macromolecule catabolic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

MYCS2:RIPA

involved_in

GO:0016998: cell wall macromolecule catabolic process

ECO:0000316: genetic interaction evidence used in manual assertion

UniProtKB:A0R3E0

P

Seeded From UniProt

complete

MYCS2:RIPA

involved_in

GO:0009253: peptidoglycan catabolic process

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

MYCS2:RIPA

involved_in

GO:0009253: peptidoglycan catabolic process

ECO:0000316: genetic interaction evidence used in manual assertion

UniProtKB:A0R3E0

P

Seeded From UniProt

complete

MYCS2:RIPA

involved_in

GO:0008360: regulation of cell shape

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

MYCS2:RIPA

involved_in

GO:0051301: cell division

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

MYCS2:RIPA

involved_in

GO:0071554: cell wall organization or biogenesis

ECO:0000316: genetic interaction evidence used in manual assertion

UniProtKB:O53168

P

Seeded From UniProt

complete

MYCS2:RPFB

involved_in

GO:0016998: cell wall macromolecule catabolic process

ECO:0000316: genetic interaction evidence used in manual assertion

UniProtKB:A0QX22

P

Seeded From UniProt

complete

MYCS2:RPFB

involved_in

GO:0009253: peptidoglycan catabolic process

ECO:0000316: genetic interaction evidence used in manual assertion

UniProtKB:A0QX22

P

Seeded From UniProt

complete

MYCTU:RIPA

involved_in

GO:0071554: cell wall organization or biogenesis

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

Notes

See also

References

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