Detailed information NTG0000119

Experimentally validated

Overview

• Name: addA
• Type: Machinery gene
• Locus tag: BSU_10630
• Genome accession: NC_000964
• Coordinates: 1139807..1143505 (+)
• Length: 1232 a.a.
• NCBI ID: NP_388944.2
• Uniprot ID: P23478
• Organism: Bacillus subtilis subsp. subtilis str. 168
• Function: homologous recombination; plasmid transformation Homologous recombination

Function

AddA, along with AddB, forms a heterodimer that functions as a nuclease, processing incoming double-stranded DNA (dsDNA) to generate single-stranded DNA (ssDNA) intermediates. This processing is crucial for the subsequent steps of homologous recombination.

Genomic Context

Location: 1134807..1148505

Locus tag	Gene name	Coordinates (strand)	Size (bp)	Protein ID	Product	Description
BSU_10600 (BSU10600)	yhjQ	1135255..1135581 (-)	327	NP_388941.1	putative metal-chelating cysteine-rich protein of unknown function	-
BSU_10610 (BSU10610)	yhjR	1135699..1136136 (-)	438	NP_388942.1	putative electron carrier protein (putative sporulation gene)	-
BSU_10620 (BSU10620)	addB	1136320..1139820 (+)	3501	NP_388943.2	ATP-dependent deoxyribonuclease (subunit B)	Machinery gene
BSU_10630 (BSU10630)	addA	1139807..1143505 (+)	3699	NP_388944.2	ATP-dependent deoxyribonuclease (subunit A)	Machinery gene
BSU_10640 (BSU10640)	sbcD	1143577..1144752 (+)	1176	NP_388945.3	DNA repair exonuclease	-
BSU_10650 (BSU10650)	sbcC	1144749..1148141 (+)	3393	NP_388946.2	DNA ATP-dependent repair enzyme	-
BSU_10660 (BSU10660)	hlpB	1148155..1148457 (+)	303	NP_388947.1	HNH nuclease-like essential for DNA repair	-

Sequence

Protein

• Length: 1232 a.a.
• Molecular weight: 141087.44 Da
• Isoelectric Point: 5.0963

>NTDB_id=119 BSU_10630 NP_388944.2 1139807..1143505(+) (addA) [Bacillus subtilis subsp. subtilis str. 168]
MNIPKPADSTWTDDQWNAIVSTGQDILVAAAAGSGKTAVLVERMIRKITAEENPIDVDRLLVVTFTNASAAEMKHRIAEA
LEKELVQRPGSLHIRRQLSLLNRASISTLHSFCLQVLKKYYYLIDLDPGFRIADQTEGELIGDEVLDELFEDEYAKGEKA
FFELVDRYTTDRHDLDLQFLVKQVYEYSRSHPNPEAWLESFVHLYDVSEKSAIEELPFYQYVKEDIAMVLNGAKEKLLRA
LELTKAPGGPAPRADNFLDDLAQIDELIQHQDDFSELYKRVPAVSFKRAKAVKGDEFDPALLDEATDLRNGAKKLLEKLK
TDYFTRSPEQHLKSLAEMKPVIETLVQLVISYGKRFEAAKQEKSIIDFSDLEHYCLAILTAENDKGEREPSEAARFYQEQ
FHEVLVDEYQDTNLVQESILQLVTSGPEETGNLFMVGDVKQSIYRFRLAEPLLFLSKYKRFTESGEGTGRKIDLNKNFRS
RADILDSTNFLFKQLMGGKIGEVDYDEQAELKLGAAYPDNDETETELLLIDNAEDTDASEEAEELETVQFEAKAIAKEIR
KLISSPFKVYDGKKKTHRNIQYRDIVILLRSMPWAPQIMEELRAQGIPVYANLTSGYFEAVEVAVALSVLKVIDNPYQDI
PLASVLRSPIVGADENELSLIRLENKKAPYYEAMKDYLAAGDRSDELYQKLNTFYGHLQKWRAFSKNHSVSELIWEVYRD
TKYMDYVGGMPGGKQRQANLRVLYDRARQYESTAFRGLFRFLRFIERMQERGDDLGTARALSEQEDVVRLMTIHSSKGLE
FPVVFVAGLGRNFNMMDLNKSYLLDKELGFGTKYIHPQLRISYPTLPLIAMKKKMRRELLSEELRVLYVALTRAKEKLFL
IGSCKDHQKQLAKWQASASQTDWLLPEFDRYQARTYLDFIGPALARHRDLGDLAGVPAHADISGHPARFAVQMIHSYDLL
DDDLEERMEEKSERLEAIRRGEPVPGSFAFDEKAREQLSWTYPHQEVTQIRTKQSVSEIKRKREYEDEYSGRAPVKPADG
SILYRRPAFMMKKGLTAAEKGTAMHTVMQHIPLSHVPSIEEAEQTVHRLYEKELLTEEQKDAIDIEEIVQFFHTEIGGQL
IGAKWKDREIPFSLALPAKEIYPDAHEADEPLLVQGIIDCLYETEDGLYLLDYKSDRIEGKFQHGFEGAAPILKKRYETQ
IQLYTKAVEQIAKTKVKGCALYFFDGGHILTL

Nucleotide

• Length: 3699 bp

>NTDB_id=119 BSU_10630 NP_388944.2 1139807..1143505(+) (addA) [Bacillus subtilis subsp. subtilis str. 168]
ATGAACATTCCTAAACCGGCAGACAGCACATGGACAGATGACCAATGGAATGCCATTGTTTCAACCGGCCAGGATATTCT
TGTGGCAGCGGCTGCGGGCTCTGGTAAAACCGCTGTGCTCGTTGAACGAATGATTCGGAAAATCACCGCGGAGGAAAACC
CAATAGATGTAGACCGTCTTCTCGTTGTGACATTTACAAACGCCTCAGCGGCAGAGATGAAGCACCGAATCGCAGAAGCC
TTGGAAAAAGAGCTTGTACAGCGCCCCGGCTCGCTGCATATTAGACGCCAGCTGTCTCTTTTAAACCGGGCCAGCATTTC
GACGCTCCATTCCTTTTGCCTGCAAGTGCTGAAGAAATATTACTACTTGATCGATCTTGATCCGGGCTTTCGGATAGCTG
ATCAGACGGAAGGCGAGCTGATCGGGGATGAAGTGCTTGACGAGCTGTTTGAAGACGAATACGCAAAGGGCGAAAAGGCG
TTTTTTGAGCTTGTTGACCGCTATACGACAGACCGGCATGATCTGGATCTGCAATTTCTCGTTAAACAGGTGTACGAGTA
TTCCCGATCCCATCCCAACCCGGAGGCGTGGCTGGAAAGCTTTGTTCATTTGTATGATGTATCAGAAAAGAGCGCCATCG
AGGAGCTGCCGTTTTATCAATATGTCAAAGAAGATATTGCAATGGTGCTTAACGGGGCGAAGGAAAAGCTCTTGCGCGCG
CTTGAGCTGACGAAAGCGCCGGGCGGCCCGGCGCCGCGTGCTGACAATTTTCTTGATGATCTTGCTCAGATTGATGAACT
GATTCAGCATCAGGACGATTTCAGTGAACTATATAAGCGGGTGCCCGCCGTCTCTTTTAAGCGTGCCAAAGCAGTAAAAG
GGGATGAGTTCGATCCAGCGCTCCTTGATGAGGCGACAGATTTGAGGAACGGCGCAAAAAAACTGCTTGAAAAGCTCAAA
ACCGACTACTTCACGCGAAGTCCTGAACAGCACTTGAAAAGCCTAGCCGAGATGAAGCCTGTCATTGAAACGCTCGTACA
GCTTGTCATCAGCTATGGAAAACGATTCGAAGCTGCGAAACAGGAAAAATCAATCATCGATTTTTCGGATTTGGAGCATT
ACTGTTTAGCGATTTTGACAGCTGAGAATGACAAAGGTGAACGTGAGCCGAGCGAGGCTGCAAGGTTTTATCAGGAACAG
TTTCATGAGGTGCTCGTTGACGAATATCAGGATACCAACCTCGTGCAGGAATCGATTCTGCAGCTCGTCACAAGCGGTCC
GGAGGAGACTGGTAACCTGTTTATGGTAGGAGATGTCAAACAGTCGATTTATCGATTCAGGCTTGCGGAGCCGCTTCTTT
TCCTCTCTAAATACAAACGGTTTACAGAGAGCGGAGAAGGCACGGGGCGGAAAATCGATTTAAATAAAAATTTCCGAAGC
CGGGCTGATATTTTAGACAGCACAAACTTTTTATTTAAACAGCTGATGGGCGGCAAAATCGGTGAGGTCGATTATGACGA
GCAGGCTGAGCTGAAGCTTGGTGCAGCGTATCCGGACAATGACGAGACGGAAACAGAGCTGCTGCTGATCGACAACGCAG
AAGATACGGATGCAAGCGAGGAAGCAGAAGAGCTTGAAACGGTGCAGTTTGAGGCAAAAGCCATCGCTAAGGAAATTCGT
AAGCTGATTTCATCGCCGTTTAAGGTGTATGACGGAAAAAAGAAAACACATCGCAATATTCAATACCGAGATATCGTGAT
TTTGCTCCGTTCGATGCCGTGGGCTCCGCAAATCATGGAGGAGCTGAGAGCACAGGGCATACCGGTTTACGCCAATTTAA
CGTCAGGCTATTTTGAAGCGGTCGAAGTCGCCGTCGCGCTTTCTGTGCTGAAGGTGATTGATAATCCGTATCAGGATATA
CCGCTTGCCTCTGTGCTGCGCTCACCGATTGTCGGAGCAGATGAAAACGAGCTGTCTTTGATCCGGCTTGAAAATAAAAA
AGCGCCGTACTATGAGGCGATGAAAGACTACCTGGCTGCTGGTGACCGGAGCGATGAGCTTTATCAAAAGTTAAATACGT
TTTACGGACATCTGCAAAAATGGCGCGCGTTTTCGAAAAACCACTCAGTATCTGAGCTGATTTGGGAAGTGTACCGCGAC
ACCAAATATATGGATTATGTCGGCGGCATGCCGGGCGGAAAACAGCGCCAGGCCAATTTGCGTGTTCTTTATGACAGGGC
GCGTCAATATGAATCAACGGCATTTCGCGGCTTGTTCCGTTTCCTGCGGTTTATCGAACGCATGCAGGAGCGGGGCGATG
ATCTTGGTACGGCGAGAGCGCTCAGCGAGCAGGAGGATGTTGTCCGCTTAATGACGATCCACAGCAGCAAAGGGCTCGAA
TTTCCAGTCGTGTTTGTAGCAGGTCTCGGCCGGAATTTCAACATGATGGATTTGAACAAATCGTACCTGCTGGATAAGGA
GCTCGGATTTGGCACGAAGTATATTCATCCGCAATTACGCATCAGCTATCCGACACTTCCGCTCATTGCGATGAAGAAAA
AAATGCGCAGGGAGCTGCTGTCAGAGGAATTGCGTGTGCTCTATGTTGCATTAACGAGAGCGAAGGAAAAGCTGTTTCTG
ATTGGCTCATGTAAGGATCATCAGAAACAGCTTGCAAAATGGCAGGCATCCGCGTCCCAAACTGATTGGCTTCTGCCGGA
ATTTGACCGCTATCAGGCGAGAACGTATCTAGATTTCATTGGGCCGGCTCTTGCCAGGCACAGAGACTTGGGGGATTTGG
CTGGTGTGCCAGCACACGCTGACATCTCAGGTCACCCGGCTCGTTTTGCCGTTCAAATGATCCATTCCTATGATTTGCTT
GATGATGATCTGGAAGAAAGAATGGAAGAAAAAAGCGAGCGCCTAGAAGCGATCCGCCGAGGTGAACCAGTTCCCGGCTC
GTTTGCGTTTGATGAAAAAGCCCGCGAGCAGCTGAGCTGGACCTACCCGCATCAAGAAGTGACGCAAATTCGGACAAAGC
AATCAGTTTCTGAGATCAAGAGAAAAAGAGAGTACGAGGATGAATACAGCGGCAGGGCCCCTGTAAAACCGGCTGATGGA
AGCATTCTGTACAGACGTCCCGCTTTTATGATGAAAAAAGGCCTGACAGCGGCAGAGAAAGGGACTGCCATGCATACGGT
TATGCAGCATATCCCGCTGTCACATGTGCCGTCGATAGAAGAAGCTGAGCAGACGGTTCACAGGCTTTATGAAAAAGAGC
TTCTCACTGAAGAACAAAAAGACGCTATTGATATAGAAGAAATCGTGCAATTTTTCCATACAGAAATCGGCGGACAGCTG
ATCGGTGCTAAGTGGAAGGACCGGGAAATACCATTCAGCTTAGCGCTTCCGGCCAAGGAGATCTATCCTGATGCACACGA
GGCAGATGAGCCGCTTTTAGTGCAGGGTATTATTGACTGTCTCTATGAAACTGAGGACGGATTATATCTATTGGATTATA
AGTCGGACCGGATTGAGGGCAAATTCCAGCATGGATTTGAAGGAGCGGCCCCGATCTTGAAGAAACGATATGAAACGCAA
ATTCAGCTGTACACGAAGGCAGTGGAGCAAATTGCAAAAACAAAGGTAAAGGGATGTGCGCTTTATTTCTTTGACGGAGG
GCACATTCTGACATTATAG

Domains

Predicted by InterproScan.

• PD-(D/E)XK nuclease superfamily (PDDEXK_1) (1053-1223)

• UvrD-like helicase C-terminal domain (UvrD_C) (472-882)

• UvrD/REP helicase N-terminal domain (UvrD-helicase) (11-461)

3D structure

• PDB: 3U44
• PDB: 3U4Q
• PDB: 4CEH
• PDB: 4CEI
• PDB: 4CEJ

References

• [1]: Neville S Gilhooly et al. (2016) Chi hotspots trigger a conformational change in the helicase-like domain of AddAB to activate homologous recombination. Nucleic Acids Research 44(6):2727-41. [PMID: 26762979]
• [2]: Wojciech W Krajewski et al. (2014) Structural basis for translocation by AddAB helicase-nuclease and its arrest at χ sites. Nature 508(7496):416-9. [PMID: 24670664]
• [3]: Kayarat Saikrishnan et al. (2012) Insights into Chi recognition from the structure of an AddAB-type helicase-nuclease complex. The EMBO Journal 31(6):1568-78. [PMID: 22307084]
• [4]: Joseph T P Yeeles et al. (2011) The AddAB helicase-nuclease catalyses rapid and processive DNA unwinding using a single Superfamily 1A motor domain. Nucleic Acids Research 39(6):2271-85. [PMID: 21071401]
• [5]: Gareth A Cromie (2009) Phylogenetic ubiquity and shuffling of the bacterial RecBCD and AddAB recombination complexes. Journal of Bacteriology 191(16):5076-84. [PMID: 19542287]
• [6]: J Kooistra et al. (1997) A conserved helicase motif of the AddA subunit of the Bacillus subtilis ATP-dependent nuclease (AddAB) is essential for DNA repair and recombination. Molecular Microbiology 23(1):137-49. [PMID: 9004227]
• [7]: B J Haijema et al. (1996) The C terminus of the AddA subunit of the Bacillus subtilis ATP-dependent DNase is required for the ATP-dependent exonuclease activity but not for the helicase activity. Journal of Bacteriology 178(17):5086-91. [PMID: 8752323]
• [8]: B J Haijema et al. (1995) Expression of the ATP-dependent deoxyribonuclease of Bacillus subtilis is under competence-mediated control. Molecular Microbiology 15(2):203-11. [PMID: 7746142]
• [9]: J Kooistra et al. (1993) The Bacillus subtilis addAB genes are fully functional in Escherichia coli. Molecular Microbiology 7(6):915-23. [PMID: 8387145]
• [10]: J C Alonso et al. (1993) Genetic recombination in Bacillus subtilis 168: effect of recN, recF, recH and addAB mutations on DNA repair and recombination. Molecular & General Genetics : MGG 239(1-2):129-36. [PMID: 8510642]