RELA_STREQ
ID RELA_STREQ Reviewed; 739 AA.
AC Q54089;
DT 01-NOV-1997, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1997, sequence version 1.
DT 03-AUG-2022, entry version 124.
DE RecName: Full=Bifunctional (p)ppGpp synthase/hydrolase RelA;
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 I;
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=relA; Synonyms=rel;
OS Streptococcus dysgalactiae subsp. equisimilis (Streptococcus equisimilis).
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Streptococcaceae;
OC Streptococcus.
OX NCBI_TaxID=119602;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=H46A;
RX PubMed=8232196; DOI=10.1007/bf00280210;
RA Mechold U., Steiner K., Vettermann S., Malke H.;
RT "Genetic organization of the streptokinase region of the Streptococcus
RT equisimilis H46A chromosome.";
RL Mol. Gen. Genet. 241:129-140(1993).
RN [2]
RP BIOPHYSICOCHEMICAL PROPERTIES, AND ACTIVITY REGULATION.
RX PubMed=12003927; DOI=10.1128/jb.184.11.2878-2888.2002;
RA Mechold U., Murphy H., Brown L., Cashel M.;
RT "Intramolecular regulation of the opposing (p)ppGpp catalytic activities of
RT Rel(Seq), the Rel/Spo enzyme from Streptococcus equisimilis.";
RL J. Bacteriol. 184:2878-2888(2002).
RN [3]
RP X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS) OF 1-385 BOUND TO
RP GUANOSINE-NUCLEOTIDE DERIVATIVES, HYDROLASE AND SYNTHASE ACTIVITIES,
RP COFACTOR, ACTIVITY REGULATION, AND MUTAGENESIS OF ARG-44; ASP-78; GLU-81;
RP ASP-82; THR-151; ASP-264 AND GLU-323.
RC STRAIN=H46A;
RX PubMed=15066282; DOI=10.1016/s0092-8674(04)00260-0;
RA Hogg T., Mechold U., Malke H., Cashel M., Hilgenfeld R.;
RT "Conformational antagonism between opposing active sites in a bifunctional
RT RelA/SpoT homolog modulates (p)ppGpp metabolism during the stringent
RT response.";
RL Cell 117:57-68(2004).
CC -!- FUNCTION: In eubacteria ppGpp (guanosine 3'-diphosphate 5'-diphosphate)
CC is a mediator of the stringent response that coordinates a variety of
CC cellular activities in response to changes in nutritional abundance.
CC This enzyme catalyzes both the formation of pppGpp which is then
CC hydrolyzed to form ppGpp, and the hydrolysis of ppGpp. The enzyme does
CC not simultaneously display both synthase and hydrolase activities. In
CC the structure of residues 1-385 there are 2 conformations seen, the
CC hydrolase-OFF/synthase-ON and hydrolase-ON/synthase-OFF, suggesting
CC there is ligand-induced signal transmission between the 2 active sites.
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=Mg(2+); Xref=ChEBI:CHEBI:18420; Evidence={ECO:0000305};
CC Note=Binds 1 Mg(2+) ion per subunit. {ECO:0000305};
CC -!- COFACTOR:
CC Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC Evidence={ECO:0000269|PubMed:15066282};
CC Note=Binds 1 Mn(2+) ion per subunit. {ECO:0000269|PubMed:15066282};
CC -!- ACTIVITY REGULATION: Alpha-beta methylenyl ATP, an ATP-analog inhibitor
CC of the synthase activity also reduces the hydrolase activity about 4-
CC fold. {ECO:0000269|PubMed:12003927, ECO:0000269|PubMed:15066282}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=2 mM for GTP {ECO:0000269|PubMed:12003927};
CC KM=5 mM for ATP {ECO:0000269|PubMed:12003927};
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 -!- DOMAIN: Based on a random mutagenesis study of the catalytic fragment
CC (residues 1-385), the (p)ppGpp phosphohydrolase domain seems to
CC encompass approximately the first 225 residues, while the (p)ppGpp
CC synthase domain seems to be found between residues 226 and 385.
CC -!- SIMILARITY: Belongs to the RelA/SpoT family. {ECO:0000305}.
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DR EMBL; X72832; CAA51353.1; -; Genomic_DNA.
DR PIR; S39975; S39975.
DR PDB; 1VJ7; X-ray; 2.10 A; A/B=1-385.
DR PDBsum; 1VJ7; -.
DR AlphaFoldDB; Q54089; -.
DR SMR; Q54089; -.
DR ChEMBL; CHEMBL1163118; -.
DR DrugBank; DB02836; Guanosine 5'-diphosphate 2':3'-cyclic monophosphate.
DR DrugBank; DB04315; Guanosine-5'-Diphosphate.
DR SABIO-RK; Q54089; -.
DR UniPathway; UPA00908; UER00884.
DR UniPathway; UPA00908; UER00886.
DR EvolutionaryTrace; Q54089; -.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR GO; GO:0005525; F:GTP binding; IEA:UniProtKB-KW.
DR GO; GO:0008728; F:GTP diphosphokinase activity; IEA:UniProtKB-EC.
DR GO; GO:0008893; F:guanosine-3',5'-bis(diphosphate) 3'-diphosphatase activity; IEA:UniProtKB-EC.
DR GO; GO:0016301; F:kinase activity; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0015970; P:guanosine tetraphosphate biosynthetic process; IEA:UniProtKB-UniPathway.
DR GO; GO:0016310; P:phosphorylation; IEA:UniProtKB-KW.
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 3D-structure; ATP-binding; GTP-binding; Hydrolase; Kinase; Magnesium;
KW Manganese; Metal-binding; Nucleotide-binding; Transferase.
FT CHAIN 1..739
FT /note="Bifunctional (p)ppGpp synthase/hydrolase RelA"
FT /id="PRO_0000166565"
FT DOMAIN 50..149
FT /note="HD"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01175"
FT DOMAIN 393..454
FT /note="TGS"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01228"
FT DOMAIN 664..739
FT /note="ACT"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01007"
FT ACT_SITE 81
FT /note="Nucleophile, for hydrolase activity"
FT /evidence="ECO:0000255"
FT ACT_SITE 82
FT /note="Nucleophile, for hydrolase activity"
FT /evidence="ECO:0000255"
FT BINDING 53
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT BINDING 77
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT BINDING 144
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT BINDING 264
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000255"
FT MUTAGEN 44
FT /note="R->Q: No hydrolase activity."
FT /evidence="ECO:0000269|PubMed:15066282"
FT MUTAGEN 78
FT /note="D->A: No hydrolase activity."
FT /evidence="ECO:0000269|PubMed:15066282"
FT MUTAGEN 81
FT /note="E->G: No hydrolase activity."
FT /evidence="ECO:0000269|PubMed:15066282"
FT MUTAGEN 82
FT /note="D->V: No hydrolase activity."
FT /evidence="ECO:0000269|PubMed:15066282"
FT MUTAGEN 151
FT /note="T->A,P: No hydrolase activity."
FT /evidence="ECO:0000269|PubMed:15066282"
FT MUTAGEN 264
FT /note="D->G: No synthase activity."
FT /evidence="ECO:0000269|PubMed:15066282"
FT MUTAGEN 323
FT /note="E->Q: No synthase activity."
FT /evidence="ECO:0000269|PubMed:15066282"
FT HELIX 9..19
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 22..38
FT /evidence="ECO:0007829|PDB:1VJ7"
FT TURN 39..41
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 52..63
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 68..76
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 79..82
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 87..94
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 96..108
FT /evidence="ECO:0007829|PDB:1VJ7"
FT TURN 128..130
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 135..150
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 160..169
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 171..177
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 181..195
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 197..209
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 211..230
FT /evidence="ECO:0007829|PDB:1VJ7"
FT TURN 231..233
FT /evidence="ECO:0007829|PDB:1VJ7"
FT STRAND 237..240
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 245..255
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 256..258
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 263..265
FT /evidence="ECO:0007829|PDB:1VJ7"
FT STRAND 267..274
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 275..288
FT /evidence="ECO:0007829|PDB:1VJ7"
FT TURN 299..301
FT /evidence="ECO:0007829|PDB:1VJ7"
FT STRAND 311..316
FT /evidence="ECO:0007829|PDB:1VJ7"
FT STRAND 318..328
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 329..337
FT /evidence="ECO:0007829|PDB:1VJ7"
FT HELIX 364..370
FT /evidence="ECO:0007829|PDB:1VJ7"
SQ SEQUENCE 739 AA; 83913 MW; E65CBEF99103D171 CRC64;
MAKEINLTGE EVVALAAKYM NETDAAFVKK ALDYATAAHF YQVRKSGEPY IVHPIQVAGI
LADLHLDAVT VACGFLHDVV EDTDITLDNI EFDFGKDVRD IVDGVTKLGK VEYKSHEEQL
AENHRKMLMA MSKDIRVILV KLADRLHNMR TLKHLRKDKQ ERISRETMEI YAPLAHRLGI
SRIKWELEDL AFRYLNETEF YKISHMMNEK RREREALVDD IVTKIKSYTT EQGLFGDVYG
RPKHIYSIYR KMRDKKKRFD QIFDLIAIRC VMETQSDVYA MVGYIHELWR PMPGRFKDYI
AAPKANGYQS IHTTVYGPKG PIEIQIRTKE MHQVAEYGVA AHWAYKKGVR GKVNQAEQKV
GMNWIKELVE LQDASNGDAV DFVDSVKEDI FSERIYVFTP TGAVQELPKD SGPIDFAYAI
HTQVGEKAIG AKVNGRMVPL TAKLKTGDVV EIVTNPNSFG PSRDWIKLVK TNKARNKIRQ
FFKNQDKELS VNKGRDMLVS YFQEQGYVAN KYLDKKRIEA ILPKVSVKSE ESLYAAVGFG
DISPVSVFNK LTEKERREEE RAKAKAEAEE LVNGGEIKHE NKDVLKVRSE NGVIIQGASG
LLMRIAKCCN PVPGDPIEGY ITKGRGIAIH RADCNNIKSQ DGYQERLIEV EWDLDNSSKD
YQAEIDIYGL NRRGLLNDVL QILSNSTKSI STVNAQPTKD MKFANIHVSF GIPNLTHLTT
VVEKIKAVPD VYSVKRTNG
