disA

168

DNA integrity scanning protein, diadenylate cyclase, delays [wiki|sporulation] in the case of chromosome damage, the DisA-dependent checkpoint arrests [wiki|DNA replication] during B. subtilis spore outgrowth until the germinating spores genome is free of damage

locus: BSU_00880

Molecular weight: 40.58 kDa

pI: 5.57

Protein length: 360 aa

Gene length: 1083 bp

function: control of [wiki|sporulation] initiation

product: DNA integrity scanning protein, has diadenylate cyclase activity

essential: no

ec: 2.7.7.85

synonyms: disA, yacK

List of homologs in different organisms, belongs to COG1623 (Galperin et al., 2021)

Gene

Coordinates: 107,476  108,558

Phenotypes of a mutant

• 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]
• reduced spore survival after infrared exposure [pubmed|28961460]
• altered morphology on MSgg medium [pubmed|29588402]
• plant root colonization defect [pubmed|29588402]

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The protein

Catalyzed reaction/ biological activity

• synthesis of c-di-AMP from two molecules of ATP [Pubmed|23192352,21566650]
• binds to abasic DNA lesions [pubmed|37250769]
• binds [protein|A44D4677FB70BE8F554BF1001A500F817C7DA95F|recA] to inhibit its activity [pubmed|30916351]
• limits [protein|24B1C2A663A5B7C45940272595868F9F05A04A7B|recG] activities at stalled or reversed replication forks [pubmed|34073022]
• limits [protein|7EBC90946E7167A3B0212193873855CAE0EFF7D4|ruvA]-[protein|8DFD57B73FE5D49BB547E050182A96CE6E8BFF77|ruvB]-mediated fork remodeling and [protein|ED1E2011C7E43A8B7E9FD9A471BC11B9DEA73177|recU]-mediated HJ cleavage to provide time for damage removal and replication restart in order to preserve genome integrity [pubmed|34768753]

Protein family

• DisA family (single member, according to UniProt)

[wiki|Domains]

• contains a [wiki|DAC domain] for the synthesis of c-di-AMP [Pubmed|18439896]
• [wiki|DAC domain] (aa 11-149) (according to UniProt)

[wiki|Cofactors]

• Mn²⁺ [pubmed|26014055]

Structure

• [PDB|3C23] [Pubmed|18439896]
• [AF|P37573]

Effectors of protein activity

• [protein|151F226370D225776F3FE7EA4901485095F1AC45|radA] is an inhibitor of [protein|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA] activity [Pubmed|23760274]
• the presence of potassium results in enhanced activity [pubmed|28420751]

[wiki|Localization]

• see a [http://download.cell.com/mmcs/journals/0092-8674/PIIS0092867406005034.mmc12.avi video] showing the movement of DisA in the cell (in real time) [Pubmed|16713562]
• forms discrete globular foci in germinating spres that colocalize with the nucleoid [Pubmed|24244006]
• forms rapidly moving focus that pauses at [protein|A44D4677FB70BE8F554BF1001A500F817C7DA95F|recA]-mediated recombination intermediates upon induction of DNA damage during spore development [pubmed|30916351]

Expression and Regulation

Operons

• genes: [gene|908DB17A39D518E84977250C55825E77FA02E391|ctsR]-[gene|CA0939064E70A97434FA450A69E779FD89880613|mcsA]-[gene|7B1B664A1AE1F641E8E7D9E2894D5C8FFFA92948|mcsB]-[gene|86A2F2F65290F4471D6FD03B694821C66C180D8A|clpC]-[gene|151F226370D225776F3FE7EA4901485095F1AC45|radA]-[gene|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA]

  description: [Pubmed|8793870]

  regulation

  • expressed during germination and spore outgrowth [Pubmed|24244006]

  regulatory mechanism

  • [protein|908DB17A39D518E84977250C55825E77FA02E391|ctsR]: repression, [pubmed|9987115,11179229,16163393,17380125], in [regulon|protein:908DB17A39D518E84977250C55825E77FA02E391|ctsR regulon]
  • [protein|2C6386E9A63F410558D168798D077DF91590F454|spx]: activation, [pubmed|30962353], in [regulon|protein:2C6386E9A63F410558D168798D077DF91590F454|spx regulon]

  sigma factors

  • [protein|081DF3EE9FA56209D648C7677188C61CE3AA8E41|sigM]: sigma factor, [Pubmed|17434969], in [regulon|protein:081DF3EE9FA56209D648C7677188C61CE3AA8E41|sigM regulon]
  • [protein|360F48D576DE950DF79C1A2677B7A35A8D8CC30C|sigA]: sigma factor, [Pubmed|8793870], in [regulon|protein:360F48D576DE950DF79C1A2677B7A35A8D8CC30C|sigA regulon]
  • [protein|580011DE5DC40EC9E7E0512791D328FAA010DCB8|sigB]: sigma factor, [pubmed|8793870] [pubmed|11544224], in [regulon|protein:580011DE5DC40EC9E7E0512791D328FAA010DCB8|sigB regulon]
  • [protein|CFF318EE6CACDEE763FBF96A2ABB46E944679AAA|sigF]: sigma factor, [Pubmed|16497325], in [regulon|protein:CFF318EE6CACDEE763FBF96A2ABB46E944679AAA|sigF regulon]

• genes: [gene|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA]

  description: [Pubmed|17434969]

  regulation

  • expressed during germination and spore outgrowth [Pubmed|24244006]

Biological materials

Mutant

• MGNA-B932 ([gene|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA]::erm), available at the [https://shigen.nig.ac.jp/bsub/resource/strainGeneDisrupted/detail/1931 NBRP B. subtilis, Japan]
• GP2142 (Δ[gene|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA]::''tet''), available in [wiki|Jörg Stülke]'s lab
• BKG2 ([gene|151F226370D225776F3FE7EA4901485095F1AC45|radA]-[gene|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA]::''spc''), available  in [wiki|Jörg Stülke]'s lab
• 1A939 ([gene|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA]::''tet''), [Pubmed|16713562], available at [http://pasture.asc.ohio-state.edu/BGSC/getdetail.cfm?bgscid=1A939&Search=1A939 BGSC]
• GP2222 ([gene|AFE2A9998219162FE32E95628B3CA9071099B61A|cdaA]::cat [gene|92EF98B872C1AD729B0BFAD46D3FA572E9C2AA32|cdaS]::ermC [gene|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA]::''tet''), available in [wiki|Jörg Stülke]'s lab, the mutant is only viable on minimal medium at low potassium concentration [pubmed|28420751]
• BKE00880 ([gene|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA]::erm  trpC2) available at [http://bgsc.org/getdetail.php?bgscid=BKE00880 BGSC],  [Pubmed|28189581], upstream reverse: _UP1_CCCTTTTTTCTCTTTTTCCA,  downstream forward: _UP4_TGATTTCGGTTAAAACCTTA
• BKK00880 ([gene|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA]::kan  trpC2) available at [http://bgsc.org/getdetail.php?bgscid=BKK00880 BGSC],  [Pubmed|28189581], upstream reverse: _UP1_CCCTTTTTTCTCTTTTTCCA,  downstream forward: _UP4_TGATTTCGGTTAAAACCTTA
• GP2715 (Δ[gene|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA]::''spc''), available in [wiki|Jörg Stülke]'s lab
• GP2752 (Δ[gene|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA]::''cat''), available in [wiki|Jörg Stülke]'s lab
• GP2782 (Δ[gene|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA]::''kan''), available in [wiki|Jörg Stülke]'s lab

Expression vectors

• IPTG inducible expression of His-[gene|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA] in ''E. coli'': pGP2563 (in [http://biochem.web.utah.edu/hill/links/pET19b.pdf pET19b]), available in [wiki|Jörg Stülke]'s lab [pubmed|23192352]

two-hybrid system

• B. pertussis adenylate cyclase-based bacterial two hybrid system ([wiki|BACTH]), available in [wiki|Fabian Commichau]'s lab

References

Reviews

• 22933559,18714086,25869574,23812326,30224435,32472931,32603625,37022175,37954098,38052445,38551349

Original Publications

• 24244006,8793870,9987115,16713562,23760274,11544224,17434969,18439896,16713555,12493822,21566650,23192352,23608499,25616256,26014055,27150552,28420751,28511132,28961460,29536659,29588402,30877841,30916351,33510358,34073022,34768753,34880841,36613897,37250769