RELA_MYCTU
ID RELA_MYCTU Reviewed; 738 AA.
AC P9WHG9; L0TBP7; P66014; Q50638;
DT 16-APR-2014, integrated into UniProtKB/Swiss-Prot.
DT 16-APR-2014, sequence version 1.
DT 03-AUG-2022, entry version 49.
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 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=relA; OrderedLocusNames=Rv2583c; ORFNames=MTCY227.18;
OS Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv).
OC Bacteria; Actinobacteria; Corynebacteriales; Mycobacteriaceae;
OC Mycobacterium; Mycobacterium tuberculosis complex.
OX NCBI_TaxID=83332;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=9634230; DOI=10.1038/31159;
RA Cole S.T., Brosch R., Parkhill J., Garnier T., Churcher C.M., Harris D.E.,
RA Gordon S.V., Eiglmeier K., Gas S., Barry C.E. III, Tekaia F., Badcock K.,
RA Basham D., Brown D., Chillingworth T., Connor R., Davies R.M., Devlin K.,
RA Feltwell T., Gentles S., Hamlin N., Holroyd S., Hornsby T., Jagels K.,
RA Krogh A., McLean J., Moule S., Murphy L.D., Oliver S., Osborne J.,
RA Quail M.A., Rajandream M.A., Rogers J., Rutter S., Seeger K., Skelton S.,
RA Squares S., Squares R., Sulston J.E., Taylor K., Whitehead S.,
RA Barrell B.G.;
RT "Deciphering the biology of Mycobacterium tuberculosis from the complete
RT genome sequence.";
RL Nature 393:537-544(1998).
RN [2]
RP PROTEIN SEQUENCE OF 306-329, FUNCTION AS A PYROPHOSPHORYLTRANSFERASE AND
RP PYROPHOSPHOHYDROLASE, CATALYTIC ACTIVITY, AND COFACTOR.
RX PubMed=10375643; DOI=10.1016/s0378-1119(99)00114-6;
RA Avarbock D., Salem J., Li L.S., Wang Z.M., Rubin H.;
RT "Cloning and characterization of a bifunctional RelA/SpoT homologue from
RT Mycobacterium tuberculosis.";
RL Gene 233:261-269(1999).
RN [3]
RP ACTIVITY REGULATION, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=10995231; DOI=10.1021/bi001256k;
RA Avarbock D., Avarbock A., Rubin H.;
RT "Differential regulation of opposing RelMtb activities by the
RT aminoacylation state of a tRNA.ribosome.mRNA.RelMtb complex.";
RL Biochemistry 39:11640-11648(2000).
RN [4]
RP FUNCTION IN REGULATION OF STRINGENT RESPONSE, AND DISRUPTION PHENOTYPE.
RX PubMed=10940033; DOI=10.1128/jb.182.17.4889-4898.2000;
RA Primm T.P., Andersen S.J., Mizrahi V., Avarbock D., Rubin H.,
RA Barry C.E. III;
RT "The stringent response of Mycobacterium tuberculosis is required for long-
RT term survival.";
RL J. Bacteriol. 182:4889-4898(2000).
RN [5]
RP MUTAGENESIS OF HIS-80; ASP-81; GLY-241; HIS-344; ASP-632 AND CYS-633, AND
RP SUBUNIT.
RX PubMed=16026164; DOI=10.1021/bi0505316;
RA Avarbock A., Avarbock D., Teh J.S., Buckstein M., Wang Z.M., Rubin H.;
RT "Functional regulation of the opposing (p)ppGpp synthetase/hydrolase
RT activities of RelMtb from Mycobacterium tuberculosis.";
RL Biochemistry 44:9913-9923(2005).
RN [6]
RP INDUCTION.
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=20529853; DOI=10.1074/jbc.m109.088872;
RA Lu L.D., Sun Q., Fan X.Y., Zhong Y., Yao Y.F., Zhao G.P.;
RT "Mycobacterial MazG is a novel NTP pyrophosphohydrolase involved in
RT oxidative stress response.";
RL J. Biol. Chem. 285:28076-28085(2010).
RN [7]
RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=21969609; DOI=10.1074/mcp.m111.011627;
RA Kelkar D.S., Kumar D., Kumar P., Balakrishnan L., Muthusamy B., Yadav A.K.,
RA Shrivastava P., Marimuthu A., Anand S., Sundaram H., Kingsbury R.,
RA Harsha H.C., Nair B., Prasad T.S., Chauhan D.S., Katoch K., Katoch V.M.,
RA Kumar P., Chaerkady R., Ramachandran S., Dash D., Pandey A.;
RT "Proteogenomic analysis of Mycobacterium tuberculosis by high resolution
RT mass spectrometry.";
RL Mol. Cell. Proteomics 10:M111.011627-M111.011627(2011).
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, as well as the hydrolysis of ppGpp. RelA is
CC probably a key factor in the pathogenesis of M.tuberculosis as it
CC regulates the intracellular concentrations of (p)ppGpp.
CC {ECO:0000269|PubMed:10375643, ECO:0000269|PubMed:10940033}.
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 Evidence={ECO:0000269|PubMed:10375643};
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 Evidence={ECO:0000269|PubMed:10375643};
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; Evidence={ECO:0000305};
CC Note=Binds 1 Mn(2+) ion per subunit. {ECO:0000305};
CC -!- ACTIVITY REGULATION: Transferase activity occurs when RelA binds to a
CC complex containing uncharged tRNA, ribosomes, and mRNA (RelA activating
CC complex or RAC). The addition of charged tRNA to this complex has the
CC opposite effect, inhibiting transferase activity and activating
CC hydrolysis activity. {ECO:0000269|PubMed:10995231}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.41 mM for ppGpp (at 30 degrees Celsius and pH 8)
CC {ECO:0000269|PubMed:10995231};
CC KM=0.48 mM for pppGpp (at 30 degrees Celsius and pH 8)
CC {ECO:0000269|PubMed:10995231};
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: Homotrimer. {ECO:0000269|PubMed:16026164}.
CC -!- INDUCTION: By MazG. {ECO:0000269|PubMed:20529853}.
CC -!- DOMAIN: Based on a mutagenesis study of the catalytic fragment
CC (residues 1-394), the (p)ppGpp phosphohydrolase domain seems to
CC encompass approximately the first 203 residues, while the (p)ppGpp
CC synthase domain seems to be found between residues 87 and 394.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene fail to synthesize ppGpp
CC in response to starvation. {ECO:0000269|PubMed:10940033}.
CC -!- SIMILARITY: Belongs to the RelA/SpoT family. {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=CCP45379.1; Type=Erroneous initiation; Note=Extended N-terminus.; Evidence={ECO:0000305};
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DR EMBL; AL123456; CCP45379.1; ALT_INIT; Genomic_DNA.
DR PIR; F70725; F70725.
DR RefSeq; NP_217099.1; NC_000962.3.
DR PDB; 5XNX; X-ray; 3.70 A; A/B/C/D=1-394.
DR PDB; 6LXG; NMR; -; A/B=660-738.
DR PDBsum; 5XNX; -.
DR PDBsum; 6LXG; -.
DR AlphaFoldDB; P9WHG9; -.
DR SASBDB; P9WHG9; -.
DR SMR; P9WHG9; -.
DR STRING; 83332.Rv2583c; -.
DR PaxDb; P9WHG9; -.
DR DNASU; 887888; -.
DR GeneID; 887888; -.
DR KEGG; mtu:Rv2583c; -.
DR TubercuList; Rv2583c; -.
DR eggNOG; COG0317; Bacteria.
DR BRENDA; 2.7.6.5; 3445.
DR SABIO-RK; P9WHG9; -.
DR UniPathway; UPA00908; UER00884.
DR UniPathway; UPA00908; UER00886.
DR Proteomes; UP000001584; Chromosome.
DR GO; GO:0009274; C:peptidoglycan-based cell wall; HDA:UniProtKB.
DR GO; GO:0005886; C:plasma membrane; HDA:MTBBASE.
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; IDA:MTBBASE.
DR GO; GO:0008893; F:guanosine-3',5'-bis(diphosphate) 3'-diphosphatase activity; IDA:MTBBASE.
DR GO; GO:0016301; F:kinase activity; IEA:UniProtKB-KW.
DR GO; GO:0030145; F:manganese ion binding; IDA:MTBBASE.
DR GO; GO:0015970; P:guanosine tetraphosphate biosynthetic process; IEA:UniProtKB-UniPathway.
DR GO; GO:0016310; P:phosphorylation; IEA:UniProtKB-KW.
DR GO; GO:0015968; P:stringent response; IMP:MTBBASE.
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; Direct protein sequencing; GTP-binding;
KW Hydrolase; Kinase; Magnesium; Manganese; Metal-binding; Nucleotide-binding;
KW Reference proteome; Transferase.
FT CHAIN 1..738
FT /note="Bifunctional (p)ppGpp synthase/hydrolase RelA"
FT /id="PRO_0000166551"
FT DOMAIN 53..150
FT /note="HD"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01175"
FT DOMAIN 398..459
FT /note="TGS"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01228"
FT DOMAIN 662..736
FT /note="ACT"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01007"
FT REGION 568..592
FT /note="Disordered"
FT /evidence="ECO:0000256|SAM:MobiDB-lite"
FT ACT_SITE 84
FT /note="Nucleophile, for hydrolase activity"
FT /evidence="ECO:0000255"
FT ACT_SITE 85
FT /note="Nucleophile, for hydrolase activity"
FT /evidence="ECO:0000255"
FT BINDING 56
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000255"
FT BINDING 80
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000255"
FT BINDING 145
FT /ligand="Mn(2+)"
FT /ligand_id="ChEBI:CHEBI:29035"
FT /evidence="ECO:0000255"
FT BINDING 265
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000250"
FT MUTAGEN 80
FT /note="H->D: Loss of hydrolytic activity."
FT /evidence="ECO:0000269|PubMed:16026164"
FT MUTAGEN 81
FT /note="D->A: Loss of hydrolytic activity."
FT /evidence="ECO:0000269|PubMed:16026164"
FT MUTAGEN 241
FT /note="G->E: Loss of ppGpp synthase activity."
FT /evidence="ECO:0000269|PubMed:16026164"
FT MUTAGEN 344
FT /note="H->Y: Loss of ppGpp synthase activity."
FT /evidence="ECO:0000269|PubMed:16026164"
FT MUTAGEN 632
FT /note="D->A: Altered association with RAC components."
FT /evidence="ECO:0000269|PubMed:16026164"
FT MUTAGEN 633
FT /note="C->A: Altered association with RAC components."
FT /evidence="ECO:0000269|PubMed:16026164"
FT STRAND 662..667
FT /evidence="ECO:0007829|PDB:6LXG"
FT HELIX 671..683
FT /evidence="ECO:0007829|PDB:6LXG"
FT STRAND 688..699
FT /evidence="ECO:0007829|PDB:6LXG"
FT STRAND 701..707
FT /evidence="ECO:0007829|PDB:6LXG"
FT HELIX 712..723
FT /evidence="ECO:0007829|PDB:6LXG"
FT STRAND 728..734
FT /evidence="ECO:0007829|PDB:6LXG"
SQ SEQUENCE 738 AA; 81827 MW; BD74EBD5BDE4E15E CRC64;
MTAQRSTTNP VLEPLVAVHR EIYPKADLSI LQRAYEVADQ RHASQLRQSG DPYITHPLAV
ANILAELGMD TTTLVAALLH DTVEDTGYTL EALTEEFGEE VGHLVDGVTK LDRVVLGSAA
EGETIRKMIT AMARDPRVLV IKVADRLHNM RTMRFLPPEK QARKARETLE VIAPLAHRLG
MASVKWELED LSFAILHPKK YEEIVRLVAG RAPSRDTYLA KVRAEIVNTL TASKIKATVE
GRPKHYWSIY QKMIVKGRDF DDIHDLVGVR ILCDEIRDCY AAVGVVHSLW QPMAGRFKDY
IAQPRYGVYQ SLHTTVVGPE GKPLEVQIRT RDMHRTAEYG IAAHWRYKEA KGRNGVLHPH
AAAEIDDMAW MRQLLDWQRE AADPGEFLES LRYDLAVQEI FVFTPKGDVI TLPTGSTPVD
FAYAVHTEVG HRCIGARVNG RLVALERKLE NGEVVEVFTS KAPNAGPSRD WQQFVVSPRA
KTKIRQWFAK ERREEALETG KDAMAREVRR GGLPLQRLVN GESMAAVARE LHYADVSALY
TAIGEGHVSA KHVVQRLLAE LGGIDQAEEE LAERSTPATM PRRPRSTDDV GVSVPGAPGV
LTKLAKCCTP VPGDVIMGFV TRGGGVSVHR TDCTNAASLQ QQAERIIEVL WAPSPSSVFL
VAIQVEALDR HRLLSDVTRA LADEKVNILS ASVTTSGDRV AISRFTFEMG DPKHLGHLLN
AVRNVEGVYD VYRVTSAA
