BSGatlas

ssbA

BSGatlas-gene-4754

BSGatlas

Description	Information
Coordinates	4198886..4199404
Genomic Size	519 bp
Name	ssbA
Outside Links	SubtiWiki
	BsubCyc
Strand	-
Type	CDS

SubtiWiki

Description	Information
Alternative Name	ssb
	ssbA
	ssbA
Category	SW 3 Information processing
	SW 3.1 Genetics
	SW 3.1.1 DNA replication
	SW 3.1.5 DNA repair/ recombination
	SW 3.1.5.6 Other proteins
	SW 6 Groups of genes
	SW 6.1 Essential genes
	SW 6.4 Phosphoproteins
	SW 6.4.1 Phosphorylation on an Arg residue
	SW 6.4.7 Phosphorylation on a Tyr residue
Description	single-strand DNA-binding protein, part of the [SW\|replisome], promoter of [[protein\|RecA ]]DNA repair center assembly, required to internalize and to recombine ssDNA with homologous resident duplex
Function	[[category\|SW 3.1.1]], [[category\|SW 3.1.5]]
Is essential?	yes
Isoelectric point	4.82
Locus Tag	BSU_40900
Molecular weight	18.5988
Name	ssbA
Product	single-strand DNA-binding protein

RefSeq

Description	Information
Alternative Locus Tag	BSU40900
Description	Evidence 1a: Function from experimental evidencesin the studied strain; PubMedId: 12682299, 14762004,16549871, 10072765, 25138221, 26001966, 28575448; Producttype f: factor
Functions	16.9: Replicate
Locus Tag	BSU_40900
Name	ssbA
Title	single-strand DNA-binding protein
Type	CDS

BsubCyc

Description	Information
Alternative Name	ssb
Citation	Cardenas PP;Carzaniga T;Zangrossi S;Briani F;Garcia-Tirado E;Deho G;Alonso JC Polynucleotide phosphorylase exonuclease and polymerase activities on single-stranded DNA ends are modulated by RecN, SsbA and RecA proteins. Nucleic Acids Res 39(21);9250-61 (2011) PUBMED: 21859751
	Cardenas PP;Gandara C;Alonso JC DNA double strand break end-processing and RecA induce RecN expression levels in Bacillus subtilis. DNA Repair (Amst) 14;1-8 (2014) PUBMED: 24373815
	Carrasco B;Serrano E;Sanchez H;Wyman C;Alonso JC Chromosomal transformation in Bacillus subtilis is a non-polar recombination reaction. Nucleic Acids Res (2016) PUBMED: 26786319
	Carrasco B;Yadav T;Serrano E;Alonso JC Bacillus subtilis RecO and SsbA are crucial for RecA-mediated recombinational DNA repair. Nucleic Acids Res 43(12);5984-97 (2015) PUBMED: 26001966
	Costes A;Lecointe F;McGovern S;Quevillon-Cheruel S;Polard P The C-Terminal Domain of the Bacterial SSB Protein Acts as a DNA Maintenance Hub at Active Chromosome Replication Forks. PLoS Genet 6(12);e1001238 (2010) PUBMED: 21170359
	Green M;Gilhooly NS;Abedeen S;Scott DJ;Dillingham MS;Soultanas P Engineering a reagentless biosensor for single-stranded DNA to measure real-time helicase activity in Bacillus. Biosens Bioelectron 61;579-86 (2014) PUBMED: 24953846
	Green M;Hatter L;Brookes E;Soultanas P;Scott DJ Defining the Intrinsically Disordered C-Terminal Domain of SSB Reveals DNA-Mediated Compaction. J Mol Biol 428(2 Pt A);357-64 (2016) PUBMED: 26707201
	Lu X;Zhang W;Shen J EMSA and single-molecule force spectroscopy study of interactions between Bacillus subtilis single-stranded DNA-binding protein and single-stranded DNA. Langmuir 27(24);15008-15 (2011) PUBMED: 22054219
	Manfredi C;Suzuki Y;Yadav T;Takeyasu K;Alonso JC RecO-mediated DNA homology search and annealing is facilitated by SsbA. Nucleic Acids Res 38(20);6920-9 (2010) PUBMED: 20581116
	Marceau AH;Bernstein DA;Walsh BW;Shapiro W;Simmons LA;Keck JL Protein interactions in genome maintenance as novel antibacterial targets. PLoS One 8(3);e58765 (2013) PUBMED: 23536821
	Sanders GM;Dallmann HG;McHenry CS Reconstitution of the B. subtilis replisome with 13 proteins including two distinct replicases. Mol Cell 37(2);273-81 (2010) PUBMED: 20122408
	Thomas J;Lee CA;Grossman AD A conserved helicase processivity factor is needed for conjugation and replication of an integrative and conjugative element. PLoS Genet 9(1);e1003198 (2013) PUBMED: 23326247
	Vujaklija D;Macek B Detecting Posttranslational Modifications of Bacterial SSB Proteins. Methods Mol Biol 922;205-18 (2012) PUBMED: 22976189
	Yadav T;Carrasco B;Hejna J;Suzuki Y;Takeyasu K;Alonso JC Bacillus subtilis DprA recruits RecA onto single-stranded DNA and mediates annealing of complementary strands coated by SsbB and SsbA. J Biol Chem 288(31);22437-50 (2013) PUBMED: 23779106
	Yadav T;Carrasco B;Myers AR;George NP;Keck JL;Alonso JC Genetic recombination in Bacillus subtilis: a division of labor between two single-strand DNA-binding proteins. Nucleic Acids Res 40(12);5546-59 (2012) PUBMED: 22373918
	Yadav T;Carrasco B;Serrano E;Alonso JC Roles of Bacillus subtilis DprA and SsbA in RecA-mediated genetic recombination. J Biol Chem 289(40);27640-52 (2014) PUBMED: 25138221
Comment	16.9: Replicate
Description	single-strand DNA-binding protein
Gene Ontology	GO:0003677 DNA binding
	GO:0003697 single-stranded DNA binding
	GO:0006260 DNA replication
	GO:0006281 DNA repair
	GO:0006974 cellular response to DNA damage stimulus
Locus Tag	BSU40900
Molecular weight	18.742
Name	ssbA

Nicolas et al. predictions

Description	Information
Expression neg. correlated with	new_2950025_2950153_c, BSU11350, BSU28810, BSU30330, BSU07070, BSU31340, new_2948977_2949052_c, new_3105800_3106064_c, BSU30110, BSU31330
Expression pos. correlated with	BSU40910, BSU40890, BSU01150, BSU01030, BSU01160, new_4199733_4199799_c, new_3035532_3035729, BSU01040, BSU01220, BSU01170
Highly expressed condition	(aero) Cells were grown in a synthetic medium (E. Härtig, A. Hartmann, M. Schätzle, A. M. Albertini, D. Jahn, Appl Environ Microbiol 72, 5260, 2006) at 37 °C. For aerobic growth, an overnight culture was used to inoculate 100 ml of the synthetic medium to a starting OD578 of 0.05. The culture was then incubated in a 500 ml baffled flask with shaking at 250 rpm [aero]. Anaerobic growth was carried out (i) in the presence of 10 mM potassium nitrate (nitrate respiration) [nit]; or (ii) in the absence of 10 mM postassium nitrate (fermentative growth) [ferm]. The procedure for anaerobic growth was: medium was inoculated to an OD578 nm of 0.1 in flasks completely filled with medium and sealed with rubber stoppers. They were shaken at 100 rpm to minimize cell aggregation. These cultures were inoculated aerobically with an aerobically grown overnight culture. Anaerobic conditions were achieved in the stoppered flasks after a short time through the consumption of residual oxygen. Cells were harvested during the exponential growth phase.
	(G150) Purified spores were obtained by growing cells in DSM medium (P. Schaeffer, J. Millet, J. P. Aubert, Proc Natl Acad Sci U S A 54, 704, Sep, 1965) at 37°C for 48 hours after which they were washed ten times in ice cold distilled waterover a period of 5 days. Purified spores were heat activated at 70°C in Tris 10 mM pH8.4 and germination was initiated by the addition of L-alanine 10 mM (A. Moir, J Bacteriol 146, 1106, Jun, 1981). After incubation for one hour at 37°C, the culture was diluted with an equal volume of 2X LBmedium and germinating cells were harvested at 135, 150 or 180 minutes after addition of L-alanine [G135, G150 and G180].
	(Glu) A 5 ml aliquot of LB medium was inoculated using frozen culture stocks. After a few hours growth at 37°C, precultures were prepared by inoculating 5 ml of M9 with this LB culture at several different dilutions usually ranging from 500- to 2000-fold. The dilution range was chosen so that one of these precultures had grown to and OD600 of 0.5 - 1.0 after overnight inculation. The chosen M9 medium precultures [at OD600 of 0.5 - 1.0] were used to inoculate 100 mL of M9 medium in 500 mL non-baffled shake flasks to an OD600 of 0.02. Filter-sterilized carbon sources were added separately to the medium M9 at following concentration: D-Glucose 3g/L[Glu], L-Malic acid 4.5g/L[Mal], L-Malic acid + D-Glucose 3 and 2g/L[M+G], D-Fructose 3g/L[Fru], D-Gluconate 4g/L[Glucon], Pyruvate 6g/L[Pyr], Glycerol 6g/L[Gly], Glutamic acid + Succinic acid 2 and 2g/L[G+S]. Where necessary, carbon source solutions were pH neutralized with 4 M NaOH prior to addition to the medium. Cells were harvested during the exponential growth phase.
	(LBGexp) Cells were grown in Luria-Bertani medium (Sigma) supplemented with glucose 0.3 % [LBG] at 37°C with vigorous shaking in flasks. Overnight cultures were diluted 2000-fold in fresh pre-warmed medium and samples were collected during the exponential [exp], transition [tran] and stationary [stat] phases of the growth cycle .
	(LPh) Cells were harvested (i) during exponential growth in high phosphate defined medium [HPh]; (ii) during exponential growth in low phosphate defined medium [LPh] (J. P. Muller, Z. An, T. Merad, I. C. Hancock, C. R. Harwood, Microbiology 143, 947, Mar, 1997);and (iii) at three hours after the outset of the phosphate-limitation induced stationary phase [LPhT].
	(M9exp) Cells were grown in M9 supplemented with glucose (0.3 %) at 37°C with vigorous shaking. The composition of the M9 minimal medium is (per liter): 8.5 g Na2HPO4.2H20, 3 g KH2PO4, 1 g NH4Cl and 0.5 g NaCl. The following solutions were individually sterilized and added (volumes per liter of medium): 1 ml 0.1 M CaCl2.2H2O, 1 ml 1 M MgSO4.7H2O, 1 ml 50 mM Fe-Citrate. Also added was 10 ml of a trace salts solution containing (per liter): 170 mg ZnCl2, 100 mg MnCl2.4H2O, 60 mg CoCl2.6H2O, 60 mg Na2MoO4.2H2O and 43 mg CuCl2.2H2O. Overnight cultures were diluted 2000-fold in pre-warmed M9 medium and samples were harvested during exponential growth [M9exp], at the transition phase [M9tran] and during stationary phase [M9stat].
	(MG+10) A culture of LB medium was inocualted from a frozen glycerol stock of B. subtilis. After few hours at 37oC when the culture was growing exponentially, this culture was used to inoculate M9 minimal medium at several different dilutions usually in the range of 500- to 2000-fold. The dilution range was chosen to ensure that at least one of these M9 precultures had reached an OD600 between 0.5 - 1.0 after overnight incubation. These precultures were then used to inoculate 2.5 L of M9 medium in a 3.1 L KLF bioreactor (Bioengineering AG, Wald, Switzerland) to a starting OD600 of 0.03 – 0.05. Condiions in the bioreactor were rigorously controlled as follows: temperature was controlled at 37 °C; the pH was maintained at exactly 7.2 by automatic titration with 2.0 M KOH and 2.0 M H2SO4, and the dissolved oxygen tension was maintained above 50%. In each nutritional shift experiment cells were grown on the single substrate until the OD600 reached 0.50, at which point the second substrate was added instantaneously (4 g/L L-malate or 3 g/L glucose). The nutrient shifts performed were from glucose to glucose+malate [GM] and from malate to malate+glucose [MG] (Buescher et al., accompanying paper). Cell growth during the course was monitored throughout the experiment by measuring OD600.
	(S0) Cells were grown in CH medium at 37°C and sporulation was induced by resuspension in warm sporulation medium as described by Sterlini and Mandelstam (J. M. Sterlini, J. Mandelstam, Biochem J 113, 29, Jun, 1969). The initiation of sporulation was designated T0, the time of resuspension. Samples were harvested at hourly intervals for 6 hours [S0 to S6] for the first set of experiments and for 8 hours [S0 to S8] for a second set of experiments.
	(T-4.40H) Anon-sporulating B. subtilis strain was grown in a modified M9 medium in batch culture (T. Hardiman, K. Lemuth, M. A. Keller, M. Reuss, M. Siemann-Herzberg, J Biotechnol 132, 359, Dec 1, 2007). Glucose was exhausted when the culture reached an OD600 of approx. 10 and this was designated T0 [T0.0H]. 7 samples were harvested at various times before glucose exhaustion [T-5.40H to T-0.40H] and 10 samples at various times after glucose exhaustion [T0.30H to T5.0H].
	(T-5.40H) Anon-sporulating B. subtilis strain was grown in a modified M9 medium in batch culture (T. Hardiman, K. Lemuth, M. A. Keller, M. Reuss, M. Siemann-Herzberg, J Biotechnol 132, 359, Dec 1, 2007). Glucose was exhausted when the culture reached an OD600 of approx. 10 and this was designated T0 [T0.0H]. 7 samples were harvested at various times before glucose exhaustion [T-5.40H to T-0.40H] and 10 samples at various times after glucose exhaustion [T0.30H to T5.0H].
Lowely expressed condition	(BC) Cultures were inoculated from frozen glycerol stocks and grown overnight in LB at 37°C. These cultures were thendiluted, plated onto LB plates, and incubated for 16 h at 37°C. Cells were harvested from plates containing individual colonies [BI] andfrom plates with confluen growth [BC].
	(Etha) Cells were grown in a synthetic medium (J. Stülke, R. Hanschke, M. Hecker, J Gen Microbiol 139, 2041, Sep, 1993) with 0.2 % glucose as carbon source (Belitsky Minimal Medium/BMM) at 37 °C with vigorous shaking. Stress was applied to exponentially growing cultures at OD500nm of 0.4. Samples were harvested before stress [BMM]; after a rapid temperature up-shift from 37 °C to 48 °C [Heat]; after a temperature down-shift from 37 °C to 18 °C [Cold]. Ethanol stress was imposed by adding ethanol to a final concentration of 4 % (v/v) and cells were harvested 10 minutes after ethanol addition [Etha].
	(LBstat) Cells were grown in Luria-Bertani medium (Sigma) [LB] at 37°C with vigorous shaking in flasks. Overnight cultures were diluted 2000-fold in fresh pre-warmed medium and samples were collected during the exponential [exp], transition [tran] and stationary [stat] phases of the growth cycle .
	(M0t90) Cells were grown in LB medium at 37°C with vigorous shaking. An exponentially growing culture (O.D.600 approx. 0.25) was divided: one culture acted as the control [no mitomycin C , M0] while mitomycin was added to the second culture to a final concentration of 40 ng/ml [mitomycin, M40]. Samples were harvested at 0, 45 and 90 minutes after mitomycin addition [t0, t45 and t90].
	(M40t90) Cells were grown in LB medium at 37°C with vigorous shaking. An exponentially growing culture (O.D.600 approx. 0.25) was divided: one culture acted as the control [no mitomycin C , M0] while mitomycin was added to the second culture to a final concentration of 40 ng/ml [mitomycin, M40]. Samples were harvested at 0, 45 and 90 minutes after mitomycin addition [t0, t45 and t90].
	(M9stat) Cells were grown in M9 supplemented with glucose (0.3 %) at 37°C with vigorous shaking. The composition of the M9 minimal medium is (per liter): 8.5 g Na2HPO4.2H20, 3 g KH2PO4, 1 g NH4Cl and 0.5 g NaCl. The following solutions were individually sterilized and added (volumes per liter of medium): 1 ml 0.1 M CaCl2.2H2O, 1 ml 1 M MgSO4.7H2O, 1 ml 50 mM Fe-Citrate. Also added was 10 ml of a trace salts solution containing (per liter): 170 mg ZnCl2, 100 mg MnCl2.4H2O, 60 mg CoCl2.6H2O, 60 mg Na2MoO4.2H2O and 43 mg CuCl2.2H2O. Overnight cultures were diluted 2000-fold in pre-warmed M9 medium and samples were harvested during exponential growth [M9exp], at the transition phase [M9tran] and during stationary phase [M9stat].
	(S5) Cells were grown in CH medium at 37°C and sporulation was induced by resuspension in warm sporulation medium as described by Sterlini and Mandelstam (J. M. Sterlini, J. Mandelstam, Biochem J 113, 29, Jun, 1969). The initiation of sporulation was designated T0, the time of resuspension. Samples were harvested at hourly intervals for 6 hours [S0 to S6] for the first set of experiments and for 8 hours [S0 to S8] for a second set of experiments.
	(S6) Cells were grown in CH medium at 37°C and sporulation was induced by resuspension in warm sporulation medium as described by Sterlini and Mandelstam (J. M. Sterlini, J. Mandelstam, Biochem J 113, 29, Jun, 1969). The initiation of sporulation was designated T0, the time of resuspension. Samples were harvested at hourly intervals for 6 hours [S0 to S6] for the first set of experiments and for 8 hours [S0 to S8] for a second set of experiments.
	(S7) Cells were grown in CH medium at 37°C and sporulation was induced by resuspension in warm sporulation medium as described by Sterlini and Mandelstam (J. M. Sterlini, J. Mandelstam, Biochem J 113, 29, Jun, 1969). The initiation of sporulation was designated T0, the time of resuspension. Samples were harvested at hourly intervals for 6 hours [S0 to S6] for the first set of experiments and for 8 hours [S0 to S8] for a second set of experiments.
	(S8) Cells were grown in CH medium at 37°C and sporulation was induced by resuspension in warm sporulation medium as described by Sterlini and Mandelstam (J. M. Sterlini, J. Mandelstam, Biochem J 113, 29, Jun, 1969). The initiation of sporulation was designated T0, the time of resuspension. Samples were harvested at hourly intervals for 6 hours [S0 to S6] for the first set of experiments and for 8 hours [S0 to S8] for a second set of experiments.
Name	ssbA

KEGG Pathways

Description	Information
Pathway	DNA replication (ko03030)
	Mismatch repair (ko03430)
	Homologous recombination (ko03440)

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