cdaA [2017-05-21 19:45:59]

diadenylate cyclase, synthesis of c-di-AMP in vegetative cells

locus: BSU01750

pI: 8.07

mw: 30.42 kDa

protein length: 273 aa

gene length: 819 bp

function: synthesis of c-di-AMP

product: diadenylate cyclase

essential: no

ec: 2.7.7.85

synonyms: ybbP,ybbQ

Genomic Context

categories

• [category|SW 2|Metabolism] → [category|SW 2.5|Nucleotide metabolism] → [category|SW 2.5.3|Metabolism of signalling nucleotides]
• [category|SW 6|Groups of genes] → [category|SW 6.2|Membrane proteins]

Gene

Coordinates: 196,213 → 197,034

Phenotypes of a mutant

• inactivation of ''[gene|AFE2A9998219162FE32E95628B3CA9071099B61A|cdaA]'' results in severe beta-lactam sensitivity [Pubmed|22211522]
• a [gene|AFE2A9998219162FE32E95628B3CA9071099B61A|cdaA] [gene|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA] double mutant or [gene|AFE2A9998219162FE32E95628B3CA9071099B61A|cdaA] [gene|92EF98B872C1AD729B0BFAD46D3FA572E9C2AA32|cdaS] [gene|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA] triple mutant is not viable on complex medium; however, the mutant grows at low potassium concentration (0.1 mM) [pubmed|28420751]

The protein

Catalyzed reaction/ biological activity

• synthesis of c-di-AMP from two molecules of ATP [Pubmed|23192352]

Paralogous protein(s)

• [protein|92EF98B872C1AD729B0BFAD46D3FA572E9C2AA32|CdaS]

[SW|Domains]

• contains a [SW|DAC domain] involved in the synthesis of c-di-AMP [Pubmed|21566650]

Effectors of protein activity

• the interaction with [protein|FB23F2AB0E2A82658009D115E5876E7AF1BDE5FD|CdaR] stimulates the diadenylate cyclase activity of [protein|AFE2A9998219162FE32E95628B3CA9071099B61A|CdaA] [Pubmed|23192352]

Structure

• [PDB|4RV7] (the [SW|DAC domain] and C-terminal domain of CdaA from ''Listeria monocytogenes'' (aa 101 - 273), 65% identity) [Pubmed|25605729]

[SW|Localization]

• cell membrane [Pubmed|26240071]

Expression and Regulation

Operons

• genes: [gene|AFE2A9998219162FE32E95628B3CA9071099B61A|cdaA]-[gene|FB23F2AB0E2A82658009D115E5876E7AF1BDE5FD|cdaR]-[gene|F022ACB2CBB8DA46B392CE1D26474093645F26DA|glmM]-[gene|4E23D2043D51C587027ABBF6335F148295E90B8F|glmS]
  description: [Pubmed|23192352]

  sigma factors

  • [protein|360F48D576DE950DF79C1A2677B7A35A8D8CC30C|SigA]: sigma factor, [Pubmed|22211522], in [regulon|360F48D576DE950DF79C1A2677B7A35A8D8CC30C|SigA regulon]

additional information

• CdaA levels are increased at increased potassium concentrations [pubmed|28420751]

Biological materials

Mutant

• GP94 (D''[gene|AFE2A9998219162FE32E95628B3CA9071099B61A|cdaA]''::''spc''), available in [SW|Jörg Stülke]'s lab [pubmed|28420751]
• GP997 (''[gene|AFE2A9998219162FE32E95628B3CA9071099B61A|cdaA]''::''cat''), available in [SW|Jörg Stülke]'s lab [pubmed|23192352]
• GP985 (''[gene|AFE2A9998219162FE32E95628B3CA9071099B61A|cdaA]''-''[gene|FB23F2AB0E2A82658009D115E5876E7AF1BDE5FD|cdaR]''::''cat''), available in [SW|Jörg Stülke]'s lab

Expression vector

• expression of native ''cdaA'' in ''B. subtilis'': pGP1960 (in [SW|pBQ200]), available in [SW|Jörg Stülke]'s lab
• expression of ''cdaA''-Strep in ''B. subtilis'' suitable for [SW|SPINE]: pGP1986 (in [SW|pGP382]), available in [SW|Jörg Stülke]'s lab
• IPTG inducible expression of ''cdaA''-Strep in ''E. coli'': pGP2564 (in [SW|pGP574]), available in [SW|Jörg Stülke]'s lab

lacZ fusion

• GP1339 (cat) based on [protein|search|pAC6], available in [SW|Jörg Stülke]'s lab

two-hybrid system

• B. pertussis adenylate cyclase-based bacterial two hybrid system ([SW|BACTH]), available in [SW|Jörg Stülke]'s lab. Respective plasmid: pGP1990.

FLAG-tag construct

• GP1381 ''cdaA-3xFLAG ermC'' (based on [SW|pGP1087]), available in [SW|Jörg Stülke]'s lab [pubmed|28420751]

References

Reviews

• 18714086,25869574,23812326

Original publications

• 12884008,21566650,23192352,22211522,25605729,25616256,26240071,26527648,26585449,28420751