BSGatlas

bslA

BSGatlas-gene-3650

BSGatlas

Description	Information
Coordinates	3187503..3188048
Genomic Size	546 bp
Name	bslA
Outside Links	SubtiWiki
	BsubCyc
Strand	+
Type	CDS

SubtiWiki

Description	Information
Alternative Name	bslA
	bslA
	sivB
	yuaB
Category	SW 3 Information processing
	SW 3.4 Regulation of gene expression
	SW 3.4.7 phosphorelay
	SW 3.4.7.2 Proteins controlling the activity of the kinases
	SW 4 Lifestyles
	SW 4.1 Exponential and early post-exponential lifestyles
	SW 4.1.2 Biofilm formation
	SW 4.1.2.3 Repellent surface layer
	SW 4.2 Sporulation
	SW 4.2.2 phosphorelay
	SW 4.2.2.2 Proteins controlling the activity of the kinases
	SW 6 Groups of genes
	SW 6.12 Secreted proteins
Description	bacterial hydrophobin, forms water-repellent surface layer of the biofilm, required for sliding, inhibitor of [[protein\|B29CAD04FB89ACC482EFCD29D50EDDA19145CAA2]] autophosphorylation, and subsequently of entry into [SW\|sporulation]
Function	[SW\|biofilm formation], control of entry into [SW\|sporulation] via the [SW\|phosphorelay]
Is essential?	no
Isoelectric point	9.99
Locus Tag	BSU_31080
Molecular weight	19.1118
Name	bslA
Product	biofilm surface layer, inhibitor of [[protein\|B29CAD04FB89ACC482EFCD29D50EDDA19145CAA2]] autophosphorylation

RefSeq

Description	Information
Alternative Locus Tag	BSU31080
Description	Evidence 1a: Function from experimental evidencesin the studied strain; PubMedId: 21097620, 21742882,22571672, 23904481, 28698374, 28701036, 28751732; Producttype s: structure
Functions	15.3: Cell adhesion
Locus Tag	BSU_31080
Name	bslA
Title	biofilm hydrophobic layer component
Type	CDS

BsubCyc

Description	Information
Alternative Name	sivB
	yuaB
Citation	Arnaouteli S;MacPhee CE;Stanley-Wall NR Just in case it rains: building a hydrophobic biofilm the Bacillus subtilis way. Curr Opin Microbiol 34;7-12 (2016) PUBMED: 27458867
	Brandani GB;Schor M;Morris R;Stanley-Wall N;MacPhee CE;Marenduzzo D;Zachariae U The Bacterial Hydrophobin BslA is a Switchable Ellipsoidal Janus Nanocolloid. Langmuir 31(42);11558-63 (2015) PUBMED: 26378478
	Bromley KM;Morris RJ;Hobley L;Brandani G;Gillespie RM;McCluskey M;Zachariae U;Marenduzzo D;Stanley-Wall NR;MacPhee CE Interfacial self-assembly of a bacterial hydrophobin. Proc Natl Acad Sci U S A 112(17);5419-24 (2015) PUBMED: 25870300
	Garti-Levi S;Eswara A;Smith Y;Fujita M;Ben-Yehuda S Novel modulators controlling entry into sporulation in Bacillus subtilis. J Bacteriol 195(7);1475-83 (2013) PUBMED: 23335417
	Hobley L;Ostrowski A;Rao FV;Bromley KM;Porter M;Prescott AR;MacPhee CE;van Aalten DM;Stanley-Wall NR BslA is a self-assembling bacterial hydrophobin that coats the Bacillus subtilis biofilm. Proc Natl Acad Sci U S A 110(33);13600-5 (2013) PUBMED: 23904481
	Kovacs AT;van Gestel J;Kuipers OP The protective layer of biofilm: a repellent function for a new class of amphiphilic proteins. Mol Microbiol 85(1);8-11 (2012) PUBMED: 22607588
	NA Correction for Hobley et al., BslA is a self-assembling bacterial hydrophobin that coats the Bacillus subtilis biofilm. Proc Natl Acad Sci U S A 112(38);E5371-5 (2015) PUBMED: 26324938
	Stanley-Wall NR;MacPhee CE Connecting the dots between bacterial biofilms and ice cream. Phys Biol 12(6);063001 (2015) PUBMED: 26685107
	Verhamme DT;Murray EJ;Stanley-Wall NR DegU and Spo0A jointly control transcription of two loci required for complex colony development by Bacillus subtilis. J Bacteriol 191(1);100-8 (2009) PUBMED: 18978066
Comment	BslA is an amphiphilic protein that contributes to the surface properties of B. subtilis biofilms by forming the hydrophobic surface layer of biofilms \|CITS: [22571672]\|. A bslA mutant has reduced complex colony structure in the 1A700 strain background \|CITS: [21097620]\| and shows loss of surface repellency and altered biofilm surface microstructure \|CITS: [22571672]\|. Both DegU~P \|CITS: [21742882]\| and Rok \|CITS: [21097620]\| appear to regulate transcription of bslA indirectly. The BslA protein sequences of B. subtilis strains 168 and NCIB3610 are 100% identical. BslA: "biofilm surface layer protein A" \|CITS: [22571672]\|
Description	protein involved in biofilm formation
Gene Ontology	GO:0016020 membrane
	GO:0016021 integral component of membrane
	GO:0044011 single-species biofilm formation on inanimate substrate
Locus Tag	BSU31080
Molecular weight	19.257
Name	bslA

Nicolas et al. predictions

Description	Information
Expression neg. correlated with	new_3263786_3264226_c, new_45224_45399_c, new_3132220_3132310_c, BSU25160, new_1151121_1151285, BSU31800, new_2596439_2596534, BSU_misc_RNA_1, BSU_misc_RNA_3, BSU_misc_RNA_6, BSU_misc_RNA_7, BSU_misc_RNA_8, BSU_misc_RNA_9, BSU_misc_RNA_10, BSU_misc_RNA_11, BSU_misc_RNA_12, BSU_misc_RNA_13, BSU_misc_RNA_14, BSU_misc_RNA_15, BSU_misc_RNA_16, BSU_misc_RNA_17, BSU_misc_RNA_18, BSU_misc_RNA_19, BSU_misc_RNA_20, BSU_misc_RNA_21, BSU_misc_RNA_23, BSU_misc_RNA_24, BSU_misc_RNA_25, BSU_misc_RNA_26, BSU_misc_RNA_27, BSU_misc_RNA_28, BSU_misc_RNA_29, BSU_misc_RNA_31, BSU_misc_RNA_32, BSU_misc_RNA_34, BSU_misc_RNA_36, BSU_misc_RNA_38, BSU_misc_RNA_39, BSU_misc_RNA_40, BSU_misc_RNA_41, BSU_misc_RNA_42, BSU_misc_RNA_44, BSU_misc_RNA_45, BSU_misc_RNA_46, BSU_misc_RNA_47, BSU_misc_RNA_48, BSU_misc_RNA_49, BSU_misc_RNA_50, BSU_misc_RNA_51, BSU_misc_RNA_52, BSU_misc_RNA_53, BSU_misc_RNA_54, BSU_misc_RNA_56, BSU_misc_RNA_59, BSU_misc_RNA_60, BSU_misc_RNA_63, BSU13820, BSU40440
Expression pos. correlated with	new_3187402_3187502, BSU25890, BSU25880, BSU18370, BSU06811, BSU26820, BSU26890, BSU02040, new_2740238_2740411, BSU03520
Highly expressed condition	(BI) 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].
	(BT) A fresh colony grown on an LB plate was used to inoculate 10 ml of LB and grown for 10 hoursat 30°C. This culture wasused to inoculate 10 ml of MSgg medium (S.S. Branda et al., J Bacteriol 186, 3970, Jun, 2004) and incubated with vigorous shaking. The cultures in MSgg were diluted to the same extent in 96 wells microtiterplates (5 μl for 1.5 ml of medium) and incubated without shaking at 30°C. Cells from the control cultures were harvested after 24 hours of incubation [BT]. Biofilms were harvested from 96 well plates after incubation for 36 hours [B36] and 60 hours [B60].
	(M9tran) 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].
	(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.
	(S2) 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-0.40H) 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-1.10H) 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-1.40H) 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-2.40H) 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].
	(T1.0H) 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	(Diami) Cells were grown in LB medium at 37°C. At OD540 of 0.3, the culture were divided into four subcultures and diamide 0.6 mM [Diami], paraquat 0.4 mM [Paraq], H2O2 0.1mM [H2O2] or no oxidative drug [Oxctl] were added to the medium. Samples were taken 10 minutes after addition
	(Glucon) 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.
	(GM-0.1) 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.
	(GM+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.
	(GM+5) 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.
	(H2O2) Cells were grown in LB medium at 37°C. At OD540 of 0.3, the culture were divided into four subcultures and diamide 0.6 mM [Diami], paraquat 0.4 mM [Paraq], H2O2 0.1mM [H2O2] or no oxidative drug [Oxctl] were added to the medium. Samples were taken 10 minutes after addition
	(MG-0.1) 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.
	(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	bslA

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