radA

branch migration transferase, 6-O-methylguanine-DNA methyltransferase, negative effector of [protein|2B091CCEE9E34D659771E39B1FC9050A145048AB|DisA] activity, participates in the stabilization and/or processing of holliday junction intermediates, required for repair-by-recombination

locus: BSU_00870

pI: 8.13

mw: 49.32 kDa

protein length: 458 aa

gene length: 1377 bp

function: homologous recombination, control of c-di-AMP formation, co-ordination of responses to replicative stress and genetic recombination

product: branch migration transferase, 6-O-methylguanine-DNA methyltransferase

essential: no

synonyms: sms,yacJ

Genomic Context

categories

• [category|SW 2|Metabolism] → [category|SW 2.5|Nucleotide metabolism] → [category|SW 2.5.3|Metabolism of signalling nucleotides]
• [category|SW 3|Information processing] → [category|SW 3.1|Genetics] → [category|SW 3.1.5|DNA repair/ recombination]
• [category|SW 3|Information processing] → [category|SW 3.1|Genetics] → [category|SW 3.1.5|DNA repair/ recombination] → [category|SW 3.1.5.6|Other proteins]
• [category|SW 4|Lifestyles] → [category|SW 4.3|Coping with stress] → [category|SW 4.3.1|General stress proteins (controlled by SigB)]
• [category|SW 4|Lifestyles] → [category|SW 4.3|Coping with stress] → [category|SW 4.3.4|Heat shock proteins]

Gene

Coordinates: 106,096 107,472

Phenotypes of a mutant

• reduced spore survival after infrared exposure [pubmed|28961460]
• sensitive to mitomycin C and H₂O₂-induced DNA damage [pubmed|30877841]
• impaired in transformation with chromosomal DNA [pubmed|30877841]
• no transformation with chromosomal DNA in [gene|24B1C2A663A5B7C45940272595868F9F05A04A7B|recG] mutant cells [pubmed|31350886]

The protein

Catalyzed reaction/ biological activity

• reduces the activity of [protein|2B091CCEE9E34D659771E39B1FC9050A145048AB|DisA] [pubmed|23760274]
• binds ssDNA and Holliday junctions (preferentially over dsDNA) [pubmed|30877841]
• promotes [protein|A44D4677FB70BE8F554BF1001A500F817C7DA95F|RecA]-mediated DNA strand exchange [pubmed|30877841]
• ATPase and 5' --> 3' DNA helicase activity [pubmed|30877841]
• unwinds forked DNA in the 5′→3′ direction [pubmed|31350886]

Protein family

• RecA family (together with [protein|A44D4677FB70BE8F554BF1001A500F817C7DA95F|RecA]) (according to UniProt)

[SW|Domains]

• N-terminal zinc finger (aa 10 ... 27) (according to UniProt)
• central RecA-like ATPase domain (according to UniProt)
• KNRFG motif (aa 255 ... 259) (according to UniProt)
• C-terminal Lon domain (aa 354 ... 458) (according to UniProt)

Structure

• [PDB|5LKM] (from Streptococcus pneumoniae, 63% identity) [pubmed|28561029]

[SW|Localization]

• cytoplasm (according to UniProt)

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]

  sigma factors

  • [protein|081DF3EE9FA56209D648C7677188C61CE3AA8E41|SigM]: sigma factor, [Pubmed|17434969], in [regulon|081DF3EE9FA56209D648C7677188C61CE3AA8E41|SigM regulon]
  • [protein|360F48D576DE950DF79C1A2677B7A35A8D8CC30C|SigA]: sigma factor, [Pubmed|8793870], in [regulon|360F48D576DE950DF79C1A2677B7A35A8D8CC30C|SigA regulon]
  • [protein|580011DE5DC40EC9E7E0512791D328FAA010DCB8|SigB]: sigma factor, [pubmed|8793870] [pubmed|11544224], in [regulon|580011DE5DC40EC9E7E0512791D328FAA010DCB8|SigB regulon]
  • [protein|CFF318EE6CACDEE763FBF96A2ABB46E944679AAA|SigF]: sigma factor, [Pubmed|16497325], in [regulon|CFF318EE6CACDEE763FBF96A2ABB46E944679AAA|SigF regulon]

  regulatory mechanism

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

  regulation

  • expressed during germination and spore outgrowth [Pubmed|24244006]
  • induction during diamide stress ([protein|2C6386E9A63F410558D168798D077DF91590F454|Spx]) [pubmed|30962353]

Biological materials

Mutant

• MGNA-B931 (yacJ::erm), available at the [https://shigen.nig.ac.jp/bsub/resource/strainGeneDisrupted/detail/1930 NBRP B. subtilis, Japan]
• BKG1 (''[gene|151F226370D225776F3FE7EA4901485095F1AC45|radA]''::''spc''), available in [SW|Jörg Stülke]'s lab
• BKG2 (''[gene|151F226370D225776F3FE7EA4901485095F1AC45|radA]-[gene|2B091CCEE9E34D659771E39B1FC9050A145048AB|disA]::spc''), available in [SW|Jörg Stülke]'s lab
• BKE00870 ([gene|151F226370D225776F3FE7EA4901485095F1AC45|radA]::erm trpC2) available at [http://www.bgsc.org/getdetail.php?bgscid=BKE00870 BGSC], [Pubmed|28189581], upstream reverse: _UP1_CATATATAGTGTAAGACCTC, downstream forward: _UP4_CGTACTTCATTAGGAGGATA
• BKK00870 ([gene|151F226370D225776F3FE7EA4901485095F1AC45|radA]::kan trpC2) available at [http://www.bgsc.org/getdetail.php?bgscid=BKK00870 BGSC], [Pubmed|28189581], upstream reverse: _UP1_CATATATAGTGTAAGACCTC, downstream forward: _UP4_CGTACTTCATTAGGAGGATA

Expression vectors

• pGP2693: expression in ''E. coli'', with C-terminal Strep-tag, in [SW|pGP574], available in [SW|Jörg Stülke]'s lab

References

Reviews

• 26459995

Original publications

• 8793870,9987115,15317759,23760274,11544224,25484163,11810266,28511132,28961460,28561029,26845522,30877841,31350886