SPOT_ECOLI
ID SPOT_ECOLI Reviewed; 702 AA.
AC P0AG24; P17580; Q2M7W3;
DT 20-DEC-2005, integrated into UniProtKB/Swiss-Prot.
DT 20-DEC-2005, sequence version 1.
DT 03-AUG-2022, entry version 137.
DE RecName: Full=Bifunctional (p)ppGpp synthase/hydrolase SpoT;
DE Includes:
DE RecName: Full=GTP pyrophosphokinase;
DE EC=2.7.6.5;
DE AltName: Full=(p)ppGpp synthase;
DE AltName: Full=ATP:GTP 3'-pyrophosphotransferase;
DE AltName: Full=Stringent response-like protein;
DE AltName: Full=ppGpp synthase II;
DE Includes:
DE RecName: Full=Guanosine-3',5'-bis(diphosphate) 3'-pyrophosphohydrolase;
DE EC=3.1.7.2;
DE AltName: Full=Penta-phosphate guanosine-3'-pyrophosphohydrolase;
DE Short=(ppGpp)ase;
GN Name=spoT; OrderedLocusNames=b3650, JW3625;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=2549050; DOI=10.1016/s0021-9258(18)63813-x;
RA Sarubbi E., Rudd K.E., Xiao H., Ikehara K., Kalman M., Cashel M.;
RT "Characterization of the spoT gene of Escherichia coli.";
RL J. Biol. Chem. 264:15074-15082(1989).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=7686882; DOI=10.1006/geno.1993.1230;
RA Burland V.D., Plunkett G. III, Daniels D.L., Blattner F.R.;
RT "DNA sequence and analysis of 136 kilobases of the Escherichia coli genome:
RT organizational symmetry around the origin of replication.";
RL Genomics 16:551-561(1993).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA Shao Y.;
RT "The complete genome sequence of Escherichia coli K-12.";
RL Science 277:1453-1462(1997).
RN [4]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=16738553; DOI=10.1038/msb4100049;
RA Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT and W3110.";
RL Mol. Syst. Biol. 2:E1-E5(2006).
RN [5]
RP PROBABLE (P)PPGPP SYNTHESIS ACTIVITY.
RX PubMed=2005134; DOI=10.1016/s0021-9258(19)67694-5;
RA Xiao H., Kalman M., Ikehara K., Zemel S., Glaser G., Cashel M.;
RT "Residual guanosine 3',5'-bispyrophosphate synthetic activity of relA null
RT mutants can be eliminated by spoT null mutations.";
RL J. Biol. Chem. 266:5980-5990(1991).
RN [6]
RP PPGPP SYNTHASE ACTIVITY, AND MUTAGENESIS OF ASP-293.
RC STRAIN=CF8295;
RX PubMed=12596879; DOI=10.1271/bbb.66.2735;
RA Fujita C., Maeda M., Fujii T., Iwamoto R., Ikehara K.;
RT "Identification of an indispensable amino acid for ppGpp synthesis of
RT Escherichia coli SpoT protein.";
RL Biosci. Biotechnol. Biochem. 66:2735-2738(2002).
RN [7]
RP FUNCTION IN PERSISTENCE, AND DISRUPTION PHENOTYPE.
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=14622409; DOI=10.1046/j.1365-2958.2003.03779.x;
RA Korch S.B., Henderson T.A., Hill T.M.;
RT "Characterization of the hipA7 allele of Escherichia coli and evidence that
RT high persistence is governed by (p)ppGpp synthesis.";
RL Mol. Microbiol. 50:1199-1213(2003).
RN [8]
RP INTERACTION WITH OBGE/CGTA.
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=15292126; DOI=10.1128/jb.186.16.5249-5257.2004;
RA Wout P., Pu K., Sullivan S.M., Reese V., Zhou S., Lin B., Maddock J.R.;
RT "The Escherichia coli GTPase CgtAE cofractionates with the 50S ribosomal
RT subunit and interacts with SpoT, a ppGpp synthetase/hydrolase.";
RL J. Bacteriol. 186:5249-5257(2004).
RN [9]
RP SUBCELLULAR LOCATION, LARGE RIBOSOMAL SUBUNIT ASSOCIATION, AND
RP CHARACTERIZATION UNDER AMINO ACID OR CARBON STARVATION.
RC STRAIN=K12;
RX PubMed=17616600; DOI=10.1128/jb.00315-07;
RA Jiang M., Sullivan S.M., Wout P.K., Maddock J.R.;
RT "G-protein control of the ribosome-associated stress response protein
RT SpoT.";
RL J. Bacteriol. 189:6140-6147(2007).
RN [10]
RP FUNCTION IN BIOFILM FORMATION, MUTAGENESIS OF ASP-73 AND ASP-259, AND
RP DISRUPTION PHENOTYPE.
RC STRAIN=K12 / MG1655 / AB400;
RX PubMed=19460094; DOI=10.1111/j.1365-2958.2009.06739.x;
RA Boehm A., Steiner S., Zaehringer F., Casanova A., Hamburger F., Ritz D.,
RA Keck W., Ackermann M., Schirmer T., Jenal U.;
RT "Second messenger signalling governs Escherichia coli biofilm induction
RT upon ribosomal stress.";
RL Mol. Microbiol. 72:1500-1516(2009).
RN [11]
RP FUNCTION, AND DISRUPTION PHENOTYPE.
RC STRAIN=K12 / CF7789, and K12 / MG1655 / ATCC 47076;
RX PubMed=21488981; DOI=10.1111/j.1365-2958.2011.07663.x;
RA Edwards A.N., Patterson-Fortin L.M., Vakulskas C.A., Mercante J.W.,
RA Potrykus K., Vinella D., Camacho M.I., Fields J.A., Thompson S.A.,
RA Georgellis D., Cashel M., Babitzke P., Romeo T.;
RT "Circuitry linking the Csr and stringent response global regulatory
RT systems.";
RL Mol. Microbiol. 80:1561-1580(2011).
RN [12]
RP FUNCTION IN PERSISTENCE, AND DISRUPTION PHENOTYPE.
RC STRAIN=K12 / BW25113;
RX PubMed=26051177; DOI=10.1016/j.molcel.2015.05.011;
RA Verstraeten N., Knapen W.J., Kint C.I., Liebens V., Van den Bergh B.,
RA Dewachter L., Michiels J.E., Fu Q., David C.C., Fierro A.C., Marchal K.,
RA Beirlant J., Versees W., Hofkens J., Jansen M., Fauvart M., Michiels J.;
RT "Obg and membrane depolarization are part of a microbial bet-hedging
RT strategy that leads to antibiotic tolerance.";
RL Mol. Cell 59:9-21(2015).
CC -!- FUNCTION: In eubacteria ppGpp (guanosine 3'-diphosphate 5'-diphosphate)
CC is a mediator of the stringent response which coordinates a variety of
CC cellular activities in response to changes in nutritional abundance.
CC This enzyme catalyzes both the synthesis and degradation of ppGpp. The
CC second messengers ppGpp and c-di-GMP together control biofilm formation
CC in response to translational stress; ppGpp represses biofilm formation
CC while c-di-GMP induces it. ppGpp activates transcription of CsrA-
CC antagonistic small RNAs CsrB and CsrC, which down-regulate CsrA's
CC action on translation during the stringent response (PubMed:21488981).
CC {ECO:0000269|PubMed:14622409, ECO:0000269|PubMed:19460094,
CC ECO:0000269|PubMed:21488981}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + GTP = AMP + guanosine 3'-diphosphate 5'-triphosphate;
CC Xref=Rhea:RHEA:22088, ChEBI:CHEBI:30616, ChEBI:CHEBI:37565,
CC ChEBI:CHEBI:142410, ChEBI:CHEBI:456215; EC=2.7.6.5;
CC -!- CATALYTIC ACTIVITY:
CC Reaction=guanosine 3',5'-bis(diphosphate) + H2O = diphosphate + GDP +
CC H(+); Xref=Rhea:RHEA:14253, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:33019, ChEBI:CHEBI:58189, ChEBI:CHEBI:77828; EC=3.1.7.2;
CC -!- COFACTOR:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC -!- PATHWAY: Purine metabolism; ppGpp biosynthesis; ppGpp from GDP: step
CC 1/1.
CC -!- PATHWAY: Purine metabolism; ppGpp biosynthesis; ppGpp from GTP: step
CC 1/2.
CC -!- SUBUNIT: Interacts with ObgE/CgtA (AC P42641); this association is not
CC required for pre-50S ribosome binding. Under both amino acid and carbon
CC starvation conditions about half of the protein dissociates from the
CC ribosome. {ECO:0000269|PubMed:15292126}.
CC -!- INTERACTION:
CC P0AG24; P0A6A8: acpP; NbExp=7; IntAct=EBI-543228, EBI-542566;
CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000269|PubMed:17616600}. Note=Is
CC associated with pre-50S ribosomal subunits in a salt-dependent manner.
CC -!- DISRUPTION PHENOTYPE: In the presence of wild-type relA, spoT cannot be
CC deleted because ppGpp accumulation is toxic. In the double relA/spoT
CC deletion (a ppGpp0 mutant) there is a very large increase in biofilm
CC formation (in a csrA-disrupted background) (PubMed:19460094). The
CC ppGpp0 mutant makes decreased levels of CsrA and its inhibitory small
CC RNAs (sRNA) CsrB and CsrC (PubMed:21488981). The double relA/spoT
CC deletion obviates persister cell formation in a hipA7 mutant
CC (PubMed:14622409, PubMed:26051177). {ECO:0000269|PubMed:14622409,
CC ECO:0000269|PubMed:19460094, ECO:0000269|PubMed:21488981,
CC ECO:0000269|PubMed:26051177}.
CC -!- SIMILARITY: Belongs to the RelA/SpoT family. {ECO:0000305}.
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DR EMBL; M24503; AAB00160.1; -; Genomic_DNA.
DR EMBL; L10328; AAA62003.1; -; Genomic_DNA.
DR EMBL; U00096; AAC76674.1; -; Genomic_DNA.
DR EMBL; AP009048; BAE77643.1; -; Genomic_DNA.
DR PIR; B30374; SHECGD.
DR RefSeq; NP_418107.1; NC_000913.3.
DR RefSeq; WP_000280488.1; NZ_STEB01000024.1.
DR AlphaFoldDB; P0AG24; -.
DR SMR; P0AG24; -.
DR BioGRID; 4259361; 494.
DR DIP; DIP-29378N; -.
DR IntAct; P0AG24; 60.
DR STRING; 511145.b3650; -.
DR jPOST; P0AG24; -.
DR PaxDb; P0AG24; -.
DR PRIDE; P0AG24; -.
DR EnsemblBacteria; AAC76674; AAC76674; b3650.
DR EnsemblBacteria; BAE77643; BAE77643; BAE77643.
DR GeneID; 66672455; -.
DR GeneID; 948159; -.
DR KEGG; ecj:JW3625; -.
DR KEGG; eco:b3650; -.
DR PATRIC; fig|1411691.4.peg.3056; -.
DR EchoBASE; EB0959; -.
DR eggNOG; COG0317; Bacteria.
DR HOGENOM; CLU_012300_3_0_6; -.
DR InParanoid; P0AG24; -.
DR OMA; DWISSPK; -.
DR PhylomeDB; P0AG24; -.
DR BioCyc; EcoCyc:SPOT-MON; -.
DR BioCyc; MetaCyc:SPOT-MON; -.
DR BRENDA; 2.7.6.5; 2026.
DR BRENDA; 3.1.7.2; 2026.
DR UniPathway; UPA00908; UER00884.
DR UniPathway; UPA00908; UER00886.
DR PRO; PR:P0AG24; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0008728; F:GTP diphosphokinase activity; IBA:GO_Central.
DR GO; GO:0008893; F:guanosine-3',5'-bis(diphosphate) 3'-diphosphatase activity; IDA:EcoCyc.
DR GO; GO:0016301; F:kinase activity; IEA:UniProtKB-KW.
DR GO; GO:0015970; P:guanosine tetraphosphate biosynthetic process; IEA:UniProtKB-UniPathway.
DR GO; GO:0015969; P:guanosine tetraphosphate metabolic process; IBA:GO_Central.
DR GO; GO:0015949; P:nucleobase-containing small molecule interconversion; IMP:EcoCyc.
DR GO; GO:0016310; P:phosphorylation; IEA:UniProtKB-KW.
DR GO; GO:0042594; P:response to starvation; IBA:GO_Central.
DR CDD; cd00077; HDc; 1.
DR CDD; cd05399; NT_Rel-Spo_like; 1.
DR CDD; cd01668; TGS_RSH; 1.
DR Gene3D; 3.10.20.30; -; 1.
DR Gene3D; 3.30.460.10; -; 1.
DR InterPro; IPR045865; ACT-like_dom_sf.
DR InterPro; IPR002912; ACT_dom.
DR InterPro; IPR012675; Beta-grasp_dom_sf.
DR InterPro; IPR003607; HD/PDEase_dom.
DR InterPro; IPR006674; HD_domain.
DR InterPro; IPR043519; NT_sf.
DR InterPro; IPR004811; RelA/Spo_fam.
DR InterPro; IPR045600; RelA/SpoT_AH_RIS.
DR InterPro; IPR007685; RelA_SpoT.
DR InterPro; IPR004095; TGS.
DR InterPro; IPR012676; TGS-like.
DR InterPro; IPR033655; TGS_RelA/SpoT.
DR Pfam; PF13291; ACT_4; 1.
DR Pfam; PF19296; DUF5913; 1.
DR Pfam; PF04607; RelA_SpoT; 1.
DR Pfam; PF02824; TGS; 1.
DR SMART; SM00471; HDc; 1.
DR SMART; SM00954; RelA_SpoT; 1.
DR SUPFAM; SSF55021; SSF55021; 1.
DR SUPFAM; SSF81271; SSF81271; 1.
DR SUPFAM; SSF81301; SSF81301; 1.
DR TIGRFAMs; TIGR00691; spoT_relA; 1.
DR PROSITE; PS51671; ACT; 1.
DR PROSITE; PS51831; HD; 1.
DR PROSITE; PS51880; TGS; 1.
PE 1: Evidence at protein level;
KW Cytoplasm; Hydrolase; Kinase; Manganese; Reference proteome; Transferase.
FT CHAIN 1..702
FT /note="Bifunctional (p)ppGpp synthase/hydrolase SpoT"
FT /id="PRO_0000166569"
FT DOMAIN 45..144
FT /note="HD"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01175"
FT DOMAIN 386..447
FT /note="TGS"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01228"
FT DOMAIN 628..702
FT /note="ACT"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01007"
FT MUTAGEN 73
FT /note="D->N: Obviates hydrolysis of ppGpp, moderately
FT decreases biofilm formation, loss of biofilm induction in
FT response to translation inhibitors."
FT /evidence="ECO:0000269|PubMed:19460094"
FT MUTAGEN 259
FT /note="D->N: Obviates synthesis of ppGpp, strongly
FT increases biofilm formation."
FT /evidence="ECO:0000269|PubMed:19460094"
FT MUTAGEN 293
FT /note="D->A: Unable to restore (p)ppGpp synthesis nor
FT complement a relA/spoT double disruption."
FT /evidence="ECO:0000269|PubMed:12596879"
FT MUTAGEN 294
FT /note="Y->I: Synthesizes (p)ppGpp, complements a relA/spoT
FT double disruption."
SQ SEQUENCE 702 AA; 79342 MW; A85F70F57D082EE8 CRC64;
MYLFESLNQL IQTYLPEDQI KRLRQAYLVA RDAHEGQTRS SGEPYITHPV AVACILAEMK
LDYETLMAAL LHDVIEDTPA TYQDMEQLFG KSVAELVEGV SKLDKLKFRD KKEAQAENFR
KMIMAMVQDI RVILIKLADR THNMRTLGSL RPDKRRRIAR ETLEIYSPLA HRLGIHHIKT
ELEELGFEAL YPNRYRVIKE VVKAARGNRK EMIQKILSEI EGRLQEAGIP CRVSGREKHL
YSIYCKMVLK EQRFHSIMDI YAFRVIVNDS DTCYRVLGQM HSLYKPRPGR VKDYIAIPKA
NGYQSLHTSM IGPHGVPVEV QIRTEDMDQM AEMGVAAHWA YKEHGETSTT AQIRAQRWMQ
SLLELQQSAG SSFEFIESVK SDLFPDEIYV FTPEGRIVEL PAGATPVDFA YAVHTDIGHA
CVGARVDRQP YPLSQPLTSG QTVEIITAPG ARPNAAWLNF VVSSKARAKI RQLLKNLKRD
DSVSLGRRLL NHALGGSRKL NEIPQENIQR ELDRMKLATL DDLLAEIGLG NAMSVVVAKN
LQHGDASIPP ATQSHGHLPI KGADGVLITF AKCCRPIPGD PIIAHVSPGK GLVIHHESCR
NIRGYQKEPE KFMAVEWDKE TAQEFITEIK VEMFNHQGAL ANLTAAINTT TSNIQSLNTE
EKDGRVYSAF IRLTARDRVH LANIMRKIRV MPDVIKVTRN RN
