BSGatlas

mtlR

BSGatlas-gene-506

BSGatlas

Description	Information
Coordinates	467130..469214
Genomic Size	2085 bp
Name	mtlR
Outside Links	SubtiWiki
	BsubCyc
Strand	+
Type	CDS

SubtiWiki

Description	Information
Alternative Name	mtlR
	mtlR
	ydaA
Category	SW 2 Metabolism
	SW 2.2 Carbon metabolism
	SW 2.2.2 Utilization of specific carbon sources
	SW 2.2.2.10 Utilization of mannitol
	SW 3 Information processing
	SW 3.4 Regulation of gene expression
	SW 3.4.2 Transcription factors and their control
	SW 3.4.2.3 PRD-type regulators
	SW 6 Groups of genes
	SW 6.4 Phosphoproteins
	SW 6.4.3 Phosphorylation on a Cys residue
Description	transcriptional activator of the [[gene\|5291FEF41CA547223EDF9EBAFA38D3EE550ED3F3]]-[[gene\|EE4A4336E25F5DCAC89214F55CDE203AB47AA249]]-[[gene\|4D54898D405EA25A0C74B1F02A492143D49B7B07]] operon
Function	regulation of mannitol utilization
Is essential?	no
Isoelectric point	5.31
Locus Tag	BSU_04160
Molecular weight	78.6678
Name	mtlR
Product	transcriptional activator, PRD-type

RefSeq

Description	Information
Alternative Locus Tag	BSU04160
Description	Evidence 1a: Function from experimental evidencesin the studied strain; PubMedId: 12897001, 23279188,24196428, 26159071; Product type r: regulator
Functions	16.3: Control
Locus Tag	BSU_04160
Name	mtlR
Title	transcriptional regulator, PTS-dependentactivator
Type	CDS

BsubCyc

Description	Information
Alternative Name	ydaA
Citation	Bouraoui H;Ventroux M;Noirot-Gros MF;Deutscher J;Joyet P Membrane sequestration by the EIIB domain of the mannitol permease MtlA activates the Bacillus subtilis mtl operon regulator MtlR. Mol Microbiol 87(4);789-801 (2013) PUBMED: 23279188
	Heravi KM;Altenbuchner J Regulation of the Bacillus subtilis mannitol utilization genes: promoter structure and transcriptional activation by the wild-type regulator (MtlR) and its mutants. Microbiology 160(Pt 1);91-101 (2014) PUBMED: 24196428
	Joyet P;Bouraoui H;Ake FM;Derkaoui M;Zebre AC;Cao TN;Ventroux M;Nessler S;Noirot-Gros MF;Deutscher J;Milohanic E Transcription regulators controlled by interaction with enzyme IIB components of the phosphoenolpyruvate: sugar phosphotransferase system. Biochim Biophys Acta 1834(7);1415-24 (2013) PUBMED: 23318733
	Joyet P;Derkaoui M;Bouraoui H;Deutscher J PTS-Mediated Regulation of the Transcription Activator MtlR from Different Species: Surprising Differences despite Strong Sequence Conservation. J Mol Microbiol Biotechnol 25(2-3);94-105 (2015) PUBMED: 26159071
	Joyet P;Derkaoui M;Poncet S;Deutscher J Control of Bacillus subtilis mtl operon expression by complex phosphorylation-dependent regulation of the transcriptional activator MtlR. Mol Microbiol 76(5);1279-94 (2010) PUBMED: 20444094
	Morabbi Heravi K;Wenzel M;Altenbuchner J Regulation of mtl operon promoter of Bacillus subtilis: Requirements of its use in expression vectors. Microb Cell Fact 10(1);83 (2011) PUBMED: 22014119
Comment	16.3: Control
Description	transcriptional regulator
Gene Ontology	GO:0003677 DNA binding
	GO:0005215 transporter activity
	GO:0005351 carbohydrate:proton symporter activity
	GO:0006351 transcription, DNA-templated
	GO:0006355 regulation of transcription, DNA-templated
	GO:0006810 transport
	GO:0008982 protein-N(PI)-phosphohistidine-sugar phosphotransferase activity
	GO:0009401 phosphoenolpyruvate-dependent sugar phosphotransferase system
	GO:0015992 NA
	GO:0016301 kinase activity
	GO:0016310 phosphorylation
	GO:0016740 transferase activity
	GO:0034219 carbohydrate transmembrane transport
	GO:0034220 ion transmembrane transport
Locus Tag	BSU04160
Molecular weight	78.852
Name	mtlR

Nicolas et al. predictions

Description	Information
Expression neg. correlated with	BSU23070, BSU22630, BSU20000, 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, new_698472_698611, BSU22610, BSU22620, BSU02980, new_728319_728449_c
Expression pos. correlated with	new_467042_467129, BSU35920, BSU39070, BSU35930, BSU35940, BSU28710, BSU31190, BSU35950, BSU35960, BSU31180
Highly 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].
	(dia5) Diamide was added to an exponentially growing culture (OD600 approx. 0.6) at a sub-lethal concentration(0.5 mM) and growth continued at 37°C with vigorous shaking. Samples were collected 0, 5 and 15 minutes after diamide addition [dia0, dia5 and dia15].
	(G135) 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].
	(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].
	(Gly) 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.
	(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 .
	(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.
	(Sw) Exponentially growing cells were spotted on 1 % agar LB plates and incubated at 37°C. Swarming cells were collected after 16 hours.
	(T0.30H) 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	(B36) 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].
	(BMM) 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].
	(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].
	(C30) Cellsgrown overnight on LB agar plates at 30°Cwere harvested and used to inoculate pre-warmed minimal medium at OD600 of 0.5 (D. Dubnau, R. Davidoff-Abelson, J Mol Biol 56, 209, Mar 14, 1971). After growth at 37°C with vigorous shaking, cells were diluted ten times in fresh pre-warmed minimal medium and samples were harvested after a period of 30 minutes [C30] , i.e. before maximal induction of competence, and after a period of 90 minutes [C90], i.e. when competence induction was maximal.
	(C90) Cellsgrown overnight on LB agar plates at 30°Cwere harvested and used to inoculate pre-warmed minimal medium at OD600 of 0.5 (D. Dubnau, R. Davidoff-Abelson, J Mol Biol 56, 209, Mar 14, 1971). After growth at 37°C with vigorous shaking, cells were diluted ten times in fresh pre-warmed minimal medium and samples were harvested after a period of 30 minutes [C30] , i.e. before maximal induction of competence, and after a period of 90 minutes [C90], i.e. when competence induction was maximal.
	(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].
	(GM+25) 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.
	(M+G) 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.
	(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].
	(Salt) Cells were grown in Spizizen’s minimal medium (SMM) at 37 °C with vigorous shaking. Salt was added, to a final concentration of 0.4 M to an exponentially growing culture of cells at OD500 of 0.4. Samples were harvested before [SMM] and 10 minutes after [Salt] NaCl addition.
Name	mtlR

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