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 02-NOV-2016, entry version 21. 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., RA Harris D.E., Gordon S.V., Eiglmeier K., Gas S., Barry C.E. III, RA Tekaia F., Badcock K., Basham D., Brown D., Chillingworth T., RA Connor R., Davies R.M., Devlin K., Feltwell T., Gentles S., Hamlin N., RA Holroyd S., Hornsby T., Jagels K., Krogh A., McLean J., Moule S., RA Murphy L.D., Oliver S., Osborne J., Quail M.A., Rajandream M.A., RA Rogers J., Rutter S., Seeger K., Skelton S., Squares S., Squares R., RA Sulston J.E., Taylor K., Whitehead S., Barrell B.G.; RT "Deciphering the biology of Mycobacterium tuberculosis from the RT complete genome sequence."; RL Nature 393:537-544(1998). RN [2] RP PROTEIN SEQUENCE OF 306-329, FUNCTION AS A PYROPHOSPHORYLTRANSFERASE RP AND 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 RT from Mycobacterium tuberculosis."; RL Gene 233:261-269(1999). RN [3] RP ENZYME 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 RP 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 RT long-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, RP AND 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., RA Yadav A.K., Shrivastava P., Marimuthu A., Anand S., Sundaram H., RA Kingsbury R., Harsha H.C., Nair B., Prasad T.S., Chauhan D.S., RA Katoch K., Katoch V.M., Kumar P., Chaerkady R., Ramachandran S., RA Dash D., Pandey A.; RT "Proteogenomic analysis of Mycobacterium tuberculosis by high RT resolution mass spectrometry."; RL Mol. Cell. Proteomics 10:M111.011627-M111.011627(2011). CC -!- FUNCTION: In eubacteria ppGpp (guanosine 3'-diphosphate 5-' CC diphosphate) is a mediator of the stringent response that CC coordinates a variety of cellular activities in response to CC changes in nutritional abundance. This enzyme catalyzes both the CC formation of pppGpp, which is then hydrolyzed to form ppGpp, as CC well as the hydrolysis of ppGpp. RelA is probably a key factor in CC the pathogenesis of M.tuberculosis as it regulates the CC intracellular concentrations of (p)ppGpp. CC {ECO:0000269|PubMed:10375643, ECO:0000269|PubMed:10940033}. CC -!- CATALYTIC ACTIVITY: Guanosine 3',5'-bis(diphosphate) + H(2)O = CC guanosine 5'-diphosphate + diphosphate. CC {ECO:0000269|PubMed:10375643}. CC -!- CATALYTIC ACTIVITY: ATP + GTP = AMP + guanosine 3'-diphosphate 5'- CC triphosphate. {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 -!- ENZYME REGULATION: Transferase activity occurs when RelA binds to CC a complex containing uncharged tRNA, ribosomes, and mRNA (RelA CC activating complex or RAC). The addition of charged tRNA to this CC complex has the opposite effect, inhibiting transferase activity CC and activating 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: CC step 1/1. CC -!- PATHWAY: Purine metabolism; ppGpp biosynthesis; ppGpp from GTP: CC step 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 CC ppGpp in response to starvation. {ECO:0000269|PubMed:10940033}. CC -!- SIMILARITY: Belongs to the RelA/SpoT family. {ECO:0000305}. CC -!- SIMILARITY: Contains 1 ACT domain. {ECO:0000255|PROSITE- CC ProRule:PRU01007}. CC -!- SIMILARITY: Contains 1 HD domain. {ECO:0000305}. CC -!- SEQUENCE CAUTION: CC Sequence=CCP45379.1; Type=Erroneous initiation; Note=Translation N-terminally shortened.; Evidence={ECO:0000305}; CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; AL123456; CCP45379.1; ALT_INIT; Genomic_DNA. DR PIR; F70725; F70725. DR RefSeq; NP_217099.1; NC_000962.3. DR ProteinModelPortal; P9WHG9; -. DR SMR; P9WHG9; -. DR STRING; 83332.Rv2583c; -. DR PaxDb; P9WHG9; -. DR EnsemblBacteria; CCP45379; CCP45379; Rv2583c. DR GeneID; 887888; -. DR KEGG; mtu:Rv2583c; -. DR TubercuList; Rv2583c; -. DR eggNOG; ENOG4105CWR; Bacteria. DR eggNOG; COG0317; LUCA. DR KO; K00951; -. DR BioCyc; MTBH37RV:G185E-6825-MONOMER; -. DR UniPathway; UPA00908; UER00884. DR UniPathway; UPA00908; UER00886. DR Proteomes; UP000001584; Chromosome. DR GO; GO:0005618; C:cell wall; IDA:MTBBASE. DR GO; GO:0005886; C:plasma membrane; IDA:MTBBASE. DR GO; GO:0016597; F:amino acid binding; IEA:InterPro. 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:0015969; P:guanosine tetraphosphate metabolic process; IBA:GO_Central. DR GO; GO:0009405; P:pathogenesis; IMP:MTBBASE. DR GO; GO:0015968; P:stringent response; IMP:MTBBASE. DR CDD; cd05399; NT_Rel-Spo_like; 1. DR CDD; cd01668; TGS_RelA_SpoT; 1. DR Gene3D; 3.10.20.30; -; 1. DR InterPro; IPR002912; ACT_dom. DR InterPro; IPR012675; Beta-grasp_dom. DR InterPro; IPR003607; HD/PDEase_dom. DR InterPro; IPR004811; RelA/Spo_fam. DR InterPro; IPR007685; RelA_SpoT. DR InterPro; IPR004095; TGS. DR InterPro; IPR012676; TGS-like. DR InterPro; IPR033655; TGS_RelA. DR Pfam; PF13291; ACT_4; 1. DR Pfam; PF13328; HD_4; 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; SSF81271; SSF81271; 1. DR TIGRFAMs; TIGR00691; spoT_relA; 1. DR PROSITE; PS51671; ACT; 1. PE 1: Evidence at protein level; KW ATP-binding; Complete proteome; Direct protein sequencing; KW GTP-binding; Hydrolase; Kinase; Magnesium; Manganese; Metal-binding; KW Nucleotide-binding; Reference proteome; Transferase. FT CHAIN 1 738 Bifunctional (p)ppGpp synthase/hydrolase FT RelA. FT /FTId=PRO_0000166551. FT DOMAIN 53 150 HD. FT DOMAIN 662 736 ACT. {ECO:0000255|PROSITE- FT ProRule:PRU01007}. FT ACT_SITE 84 84 Nucleophile, for hydrolase activity. FT {ECO:0000255}. FT ACT_SITE 85 85 Nucleophile, for hydrolase activity. FT {ECO:0000255}. FT METAL 56 56 Manganese. {ECO:0000255}. FT METAL 80 80 Manganese. {ECO:0000255}. FT METAL 145 145 Manganese. {ECO:0000255}. FT METAL 265 265 Magnesium. {ECO:0000250}. FT MUTAGEN 80 80 H->D: Loss of hydrolytic activity. FT {ECO:0000269|PubMed:16026164}. FT MUTAGEN 81 81 D->A: Loss of hydrolytic activity. FT {ECO:0000269|PubMed:16026164}. FT MUTAGEN 241 241 G->E: Loss of ppGpp synthase activity. FT {ECO:0000269|PubMed:16026164}. FT MUTAGEN 344 344 H->Y: Loss of ppGpp synthase activity. FT {ECO:0000269|PubMed:16026164}. FT MUTAGEN 632 632 D->A: Altered association with RAC FT components. FT {ECO:0000269|PubMed:16026164}. FT MUTAGEN 633 633 C->A: Altered association with RAC FT components. FT {ECO:0000269|PubMed:16026164}. 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 //