BSGatlas

spoIIE

BSGatlas-gene-89

BSGatlas

Description	Information
Coordinates	70538..73021
Genomic Size	2484 bp
Name	spoIIE
Outside Links	SubtiWiki
	BsubCyc
Strand	+
Type	CDS

SubtiWiki

Description	Information
Alternative Name	spoIIE
	spoIIE
	spoIIH
	spoIIK
Category	SW 3 Information processing
	SW 3.3 Protein synthesis, modification and degradation
	SW 3.3.4 Protein modification
	SW 3.3.4.4 Protein phosphatases
	SW 3.4 Regulation of gene expression
	SW 3.4.1 Sigma factors and their control
	SW 3.4.1.2 Control of sigma factors
	SW 6 Groups of genes
	SW 6.2 Membrane proteins
Description	protein serine phosphatase, septum-associated PP2C, dephosphorylation of [[protein\|3936E91C062074BFE4284B54A8CC33F35F94F5EE]]
Enzyme Classifications	EC 3.1.3.16: protein-serine/threonine phosphatase
Function	control of [[protein\|SigF ]]activity, required for normal formation of the asymmetric septum
Is essential?	no
Isoelectric point	6.14
Locus Tag	BSU_00640
Molecular weight	91.7766
Name	spoIIE
Product	protein serine phosphatase, septum-associated PP2C

RefSeq

Description	Information
Alternative Locus Tag	BSU00640
Description	Evidence 1a: Function from experimental evidencesin the studied strain; PubMedId: 10747015, 12180929,15126482, 15866939, 15978076, 28358838; Product type e :enzyme
Enzyme Classifications	EC 3.1.3.16: protein-serine/threonine phosphatase
Functions	16.13: Shape
	16.3: Control
Locus Tag	BSU_00640
Name	spoIIE
Title	SpoIIAA-phosphate serine phosphatase
Type	CDS

BsubCyc

Description	Information
Alternative Name	spoIIH
	spoIIK
Citation	Arigoni F;Duncan L;Alper S;Losick R;Stragier P SpoIIE governs the phosphorylation state of a protein regulating transcription factor sigma F during sporulation in Bacillus subtilis. Proc Natl Acad Sci U S A 93(8);3238-42 (1996) PUBMED: 8622920
	Bradshaw N;Losick R Asymmetric division triggers cell-specific gene expression through coupled capture and stabilization of a phosphatase. Elife 4 (2015) PUBMED: 26465112
	Carniol K;Ben-Yehuda S;King N;Losick R Genetic dissection of the sporulation protein SpoIIE and its role in asymmetric division in Bacillus subtilis. J Bacteriol 187(10);3511-20 (2005) PUBMED: 15866939
	Eswaramoorthy P;Winter PW;Wawrzusin P;York AG;Shroff H;Ramamurthi KS Asymmetric Division and Differential Gene Expression during a Bacterial Developmental Program Requires DivIVA. PLoS Genet 10(8);e1004526 (2014) PUBMED: 25101664
	Feucht A;Abbotts L;Errington J The cell differentiation protein SpoIIE contains a regulatory site that controls its phosphatase activity in response to asymmetric septation. Mol Microbiol 45(4);1119-30 (2002) PUBMED: 12180929
	Levdikov VM;Blagova EV;Rawlings AE;Jameson K;Tunaley J;Hart DJ;Barak I;Wilkinson AJ Structure of the phosphatase domain of the cell fate determinant SpoIIE from Bacillus subtilis. J Mol Biol 415(2);343-58 (2012) PUBMED: 22115775
	Lucet I;Feucht A;Yudkin MD;Errington J Direct interaction between the cell division protein FtsZ and the cell differentiation protein SpoIIE. EMBO J 19(7);1467-75 (2000) PUBMED: 10747015
	McBride SM;Rubio A;Wang L;Haldenwang WG Contributions of protein structure and gene position to the compartmentalization of the regulatory proteins sigma(E) and SpoIIE in sporulating Bacillus subtilis. Mol Microbiol 57(2);434-51 (2005) PUBMED: 15978076
	Muchova K;Chromikova Z;Bradshaw N;Wilkinson AJ;Barak I Morphogenic Protein RodZ Interacts with Sporulation Specific SpoIIE in Bacillus subtilis. PLoS One 11(7);e0159076 (2016) PUBMED: 27415800
	Rawlings AE;Levdikov VM;Blagova E;Colledge VL;Mas PJ;Tunaley J;Vavrova L;Wilson KS;Barak I;Hart DJ;Wilkinson AJ Expression of soluble, active fragments of the morphogenetic protein SpoIIE from Bacillus subtilis using a library-based construct screen. Protein Eng Des Sel 23(11);817-25 (2010) PUBMED: 20817757
	Searls T;Chen X;Allen S;Yudkin MD Evaluation of the kinetic properties of the sporulation protein SpoIIE of Bacillus subtilis by inclusion in a model membrane. J Bacteriol 186(10);3195-201 (2004) PUBMED: 15126482
Comment	SpoIIE is a membrane-localized serine phosphatase that establishes compartment-specific gene expression in the forespore, coupling morphological development to differential gene expression. After polar septation, SpoIIE dephosphorylates the anti-sigma factor antagonist \|FRAME: BSU23470-MONOMER "SpoIIAA"\|, which leads to the release of the sporulation-specific sigma factor \|FRAME: BSU23450-MONOMER "σ^F"\| from a complex with the anti-sigma factor \|FRAME: BSU23460-MONOMER "SpoIIAB"\|. (From \|CITS: [22115775]\|)
Description	serine phosphatase
Enzyme Classifications	EC 3.1.3.16: protein-serine/threonine phosphatase
Gene Ontology	GO:0003824 catalytic activity
	GO:0004721 phosphoprotein phosphatase activity
	GO:0004722 protein serine/threonine phosphatase activity
	GO:0005515 protein binding
	GO:0005628 prospore membrane
	GO:0005886 plasma membrane
	GO:0008152 metabolic process
	GO:0016020 membrane
	GO:0016021 integral component of membrane
	GO:0016311 dephosphorylation
	GO:0016787 hydrolase activity
	GO:0030435 sporulation resulting in formation of a cellular spore
	GO:0070262 peptidyl-serine dephosphorylation
Locus Tag	BSU00640
Molecular weight	91.968
Name	spoIIE

Nicolas et al. predictions

Description	Information
Expression neg. correlated with	BSU16330, BSU16430, BSU16440, BSU16410, BSU16390, BSU16420, BSU16400, BSU16450, BSU16460, BSU16310
Expression pos. correlated with	BSU00650, BSU19640, BSU00660, BSU27980, BSU37030, new_1605491_1605580, BSU15310, BSU15320, BSU15170, BSU27970
Highly 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].
	(B60) 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].
	(LoTm) Cells were grown in Spizizen’s minimal medium (SMM) (C. Anagnostopoulos, J. Spizizen, J Bacteriol 81, 741, May, 1961) with vigorous agitation. The control culture was grown at 37 °C [SMMPr]. For growth at high or low temperatures, pre-cultures were grown at 37 °C, diluted to an OD578nm of 0.1 and subsequently transferred to 51 °C [HiTm] and 16 °C [LoTm], respectively. For the growth at high salinity, the salinity of the medium was adjusted by adding NaCl (5 M stock solution) to produce a final concentration of 1.2 M [HiOs].
	(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].
	(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.
	(S3) 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.
	(T3.0H) 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].
	(T3.30H) 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].
	(T4.0H) 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].
	(T5.0H) 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	(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].
	(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 .
	(LBtran) 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].
	(M40t45) 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].
	(Mal) 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.
	(S1) 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	spoIIE

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