sasA

168

(p)ppGpp synthetase, small alarmone synthetase, contributes to cellular heterogeneity in protein synthesis during nutrient limitation

locus: BSU_38480

Molecular weight: 24.45 kDa

pI: 6.22

Protein length: 210 aa

Gene length: 633 bp

function: ppGpp synthesis independent from stringent response

product: (p)ppGpp synthetase

essential: no

synonyms: sasA, ywaC, relP, ipa-7d

List of homologs in different organisms, belongs to COG2357 (Galperin et al., 2021)

Gene

Coordinates: 3,949,952  3,950,584

Phenotypes of a mutant

• a [gene|CEE73284EFC0DBE8870CE0B474922DED79475A57|rel] [gene|1107A963C253FBEE8716507BD0EC0E7CD5642B12|sasA] [gene|81BCF5C08EC1A580F5E05884BA830130FC7E46E9|sasB] triple mutant requires branched chain amino acids, methionine and threonine for growth, the requirement can be suppressed by reduced expression of [gene|AC67305E4C165CC1D785E49B7CD8335346134376|guaA] or inactivation of [gene|90ACE0DECA58D3C1A55E135120F7A0C1C920571C|codY] [Pubmed|24163341]
• a [gene|CEE73284EFC0DBE8870CE0B474922DED79475A57|rel] [gene|1107A963C253FBEE8716507BD0EC0E7CD5642B12|sasA] [gene|81BCF5C08EC1A580F5E05884BA830130FC7E46E9|sasB] triple mutant acquires suppressor mutations in [gene|AC67305E4C165CC1D785E49B7CD8335346134376|guaA], [gene|AC67305E4C165CC1D785E49B7CD8335346134376|guaA], [gene|1BC994DE60A7BB35DEDD7154581A396D29AA94A7|gmk] or inactivation of [gene|90ACE0DECA58D3C1A55E135120F7A0C1C920571C|codY] [Pubmed|24682323,24163341]

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The protein

Catalyzed reaction/ biological activity

• ATP + GTP --> AMP + guanosine 3'-diphosphate 5'-triphosphate (according to UniProt)

Protein family

• RelA/SpoT family (with [protein|81BCF5C08EC1A580F5E05884BA830130FC7E46E9|sasB] and [protein|CEE73284EFC0DBE8870CE0B474922DED79475A57|rel], according to UniProt)

[wiki|Domains]

• single ppGpp synthetase domain [Pubmed|24163341]

Structure

• [PDB|6FGK] [Pubmed|29391580]
• [AF|P39583]

Paralogous protein(s)

• [protein|81BCF5C08EC1A580F5E05884BA830130FC7E46E9|sasB]

Expression and Regulation

Operons

• genes: [gene|1107A963C253FBEE8716507BD0EC0E7CD5642B12|sasA]

  description: [Pubmed|11866510]

  regulation

  • expression is reduced in a [protein|search|SigV] mutant [Pubmed|21926231]

  regulatory mechanism

  • [protein|7F340423A34CE40D1F1AA8D373F7C4B859A6496D|walR]: repression, [pubmed|31723135], in [regulon|protein:7F340423A34CE40D1F1AA8D373F7C4B859A6496D|walR regulon]

  sigma factors

  • [protein|081DF3EE9FA56209D648C7677188C61CE3AA8E41|sigM]: sigma factor, [Pubmed|18179421,11866510,18179421], in [regulon|protein:081DF3EE9FA56209D648C7677188C61CE3AA8E41|sigM regulon]
  • [protein|3D974DD2967C7F8FD4E3C2AF42617B8AE9F0296D|sigW]: sigma factor, in [regulon|protein:3D974DD2967C7F8FD4E3C2AF42617B8AE9F0296D|sigW regulon]
  • [protein|D04629D798E458AD538809BB2B150B7FA81D2C7E|sigV]: sigma factor, in [regulon|protein:D04629D798E458AD538809BB2B150B7FA81D2C7E|sigV regulon]

Biological materials

Mutant

• GP3470 (Δ[gene|1107A963C253FBEE8716507BD0EC0E7CD5642B12|sasA]::spec Δ[gene|81BCF5C08EC1A580F5E05884BA830130FC7E46E9|sasB]::phleo), available in [wiki|Jörg Stülke]'s lab
• GP3471 (Δ[gene|1107A963C253FBEE8716507BD0EC0E7CD5642B12|sasA]::spec Δ[gene|CEE73284EFC0DBE8870CE0B474922DED79475A57|rel]::cat), available in [wiki|Jörg Stülke]'s lab
• GP3424 (Δ[gene|1107A963C253FBEE8716507BD0EC0E7CD5642B12|sasA]::spec), available in [wiki|Jörg Stülke]'s lab
• MGNA-B218 (ywaC::erm), available at the [https://shigen.nig.ac.jp/bsub/resource/strainGeneDisrupted/detail/1217 NBRP B. subtilis, Japan]
• GP2066 (''[gene|1107A963C253FBEE8716507BD0EC0E7CD5642B12|sasA] [gene|81BCF5C08EC1A580F5E05884BA830130FC7E46E9|sasB] [gene|CEE73284EFC0DBE8870CE0B474922DED79475A57|rel]::mls''), available in [wiki|Jörg Stülke]'s lab
• BKE38480 ([gene|1107A963C253FBEE8716507BD0EC0E7CD5642B12|sasA]::erm  trpC2) available at [http://bgsc.org/getdetail.php?bgscid=BKE38480 BGSC],  [Pubmed|28189581], upstream reverse: _UP1_CATGTTCGTCATCTCCTTTA,  downstream forward: _UP4_TAAAAAAGACGGCACCCAAG
• BKK38480 ([gene|1107A963C253FBEE8716507BD0EC0E7CD5642B12|sasA]::kan  trpC2) available at [http://bgsc.org/getdetail.php?bgscid=BKK38480 BGSC],  [Pubmed|28189581], upstream reverse: _UP1_CATGTTCGTCATCTCCTTTA,  downstream forward: _UP4_TAAAAAAGACGGCACCCAAG

Expression vectors

• pGP3451 (N-terminal 6xHis-SUMO-tag, purification from E. coli, in pET-SUMO), available in [wiki|Jörg Stülke]'s lab
• pGP3456: expression of Strep-[gene|CEE73284EFC0DBE8870CE0B474922DED79475A57|rel] by [wiki|pGP380] in B. subtilis suitable for [wiki|SPINE], available in [wiki|Jörg Stülke]'s lab
• pGP3457: expression of [gene|CEE73284EFC0DBE8870CE0B474922DED79475A57|rel]-Strep by [wiki|pGP382] in B. subtilis suitable for [wiki|SPINE], available in [wiki|Jörg Stülke]'s lab

two-hybrid system

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

lacZ fusion

• GP3692 (based on [wiki|pAC7]), available in [wiki|Jörg Stülke]'s lab

References

Reviews

• 27149325,30980074

Original Publications

• 29391580,21926231,22950019,18670626,19447912,18067544,19477419,12207695,11866510,18179421,24163341,24489751,24687489,24682323,26460002,27875634,31723135,35594298