BSGatlas

cotH

BSGatlas-gene-4208

BSGatlas

Description	Information
Coordinates	3716009..3717097
Genomic Size	1089 bp
Name	cotH
Outside Links	SubtiWiki
	BsubCyc
Strand	-
Type	CDS

SubtiWiki

Description	Information
Alternative Name	cotH
	cotH
	ywrH
Category	SW 4 Lifestyles
	SW 4.2 Sporulation
	SW 4.2.1 Sporulation proteins
	SW 4.2.1.1 Spore coat proteins
	SW 4.2.1.1.8 Not yet assigned
Description	protein kinase, required for the assembly of several outer coat proteins
Function	protection of [[protein\|EB690FA075EAC3E298C47AED082B5FC21E8F8F46]] and [[protein\|9C59B51FC19FC83BD14272008BD441833DD1738E]] in the mother cell
Is essential?	no
Isoelectric point	5.95
Locus Tag	BSU_36060
Molecular weight	42.6537
Name	cotH
Product	protein kinase

RefSeq

Description	Information
Alternative Locus Tag	BSU36060
Description	Evidence 1a: Function from experimental evidencesin the studied strain; PubMedId: 14762006, 25115591,27185916; Product type cp: cell process
Functions	16.13: Shape
	16.5: Explore
	16.8: Protect
Locus Tag	BSU_36060
Name	cotH
Title	spore coat protein kinase
Type	CDS

BsubCyc

Description	Information
Alternative Name	ywrH
Citation	Giglio R;Fani R;Isticato R;De Felice M;Ricca E;Baccigalupi L Organization and evolution of the cotG and cotH genes of Bacillus subtilis. J Bacteriol 193(23);6664-73 (2011) PUBMED: 21984783
	Isticato R;Sirec T;Giglio R;Baccigalupi L;Rusciano G;Pesce G;Zito G;Sasso A;De Felice M;Ricca E Flexibility of the programme of spore coat formation in Bacillus subtilis: bypass of CotE requirement by over-production of CotH. PLoS One 8(9);e74949 (2013) PUBMED: 24086406
	Isticato R;Sirec T;Vecchione S;Crispino A;Saggese A;Baccigalupi L;Notomista E;Driks A;Ricca E The Direct Interaction between Two Morphogenetic Proteins Is Essential for Spore Coat Formation in Bacillus subtilis. PLoS One 10(10);e0141040 (2015) PUBMED: 26484546
	Manzourolajdad A;Arnold J Secondary structural entropy in RNA switch (Riboswitch) identification. BMC Bioinformatics 16(1);133 (2015) PUBMED: 25928324
	Nguyen KB;Sreelatha A;Durrant ES;Lopez-Garrido J;Muszewska A;Dudkiewicz M;Grynberg M;Yee S;Pogliano K;Tomchick DR;Pawlowski K;Dixon JE;Tagliabracci VS Phosphorylation of spore coat proteins by a family of atypical protein kinases. Proc Natl Acad Sci U S A 113(25);E3482-91 (2016) PUBMED: 27185916
	Plomp M;Carroll AM;Setlow P;Malkin AJ Architecture and Assembly of the Bacillus subtilis Spore Coat. PLoS One 9(9);e108560 (2014) PUBMED: 25259857
	Saggese A;Scamardella V;Sirec T;Cangiano G;Isticato R;Pane F;Amoresano A;Ricca E;Baccigalupi L Antagonistic Role of CotG and CotH on Spore Germination and Coat Formation in Bacillus subtilis. PLoS One 9(8);e104900 (2014) PUBMED: 25115591
Comment	CotH is an atypical protein kinase that phosphorylates the spore coat proteins CotB and CotG \|CITS: [27185916]\|. CotH is required for efficent spore germination \|CITS: [27185916]\|. 16.8: Protect 16.5: Explore 16.13: Shape
Description	spore coat protein kinase
Enzyme Classifications	EC 2.7.11.1: non-specific serine/threonine protein kinase
	EC 2.7.11.12: cGMP-dependent protein kinase
	EC 2.7.11.13: protein kinase C
	EC 2.7.11.17: Ca2+/calmodulin-dependent protein kinase
	EC 2.7.11.21: polo kinase
	EC 2.7.11.22: cyclin-dependent kinase
Gene Ontology	GO:0004672 protein kinase activity
	GO:0009847 spore germination
	GO:0030435 sporulation resulting in formation of a cellular spore
	GO:0031160 spore wall
	GO:0043593 endospore coat
Locus Tag	BSU36060
Molecular weight	42.813
Name	cotH

Nicolas et al. predictions

Description	Information
Expression neg. correlated with	BSU06330, BSU30400, BSU14080, new_3645297_3645378_c, new_1482096_1482247, BSU35480, BSU30390, BSU31490, BSU15000, BSU10630
Expression pos. correlated with	new_3715928_3716008_c, new_3717098_3717211_c, BSU13360, BSU10610, BSU36040, BSU05550, BSU30850, BSU22250, BSU05560, BSU32850
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].
	(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].
	(Pyr) 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.
	(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.
	(S4) 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.
	(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.
Lowely 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].
	(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.
	(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.
	(Paraq) 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
	(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.
	(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.
	(Sw) Exponentially growing cells were spotted on 1 % agar LB plates and incubated at 37°C. Swarming cells were collected after 16 hours.
	(T1.30H) 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].
	(T2.0H) 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].
Name	cotH

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