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Escherichia coli K-12 substr. MG1655 Enzyme: sulfate adenylyltransferase

Subunit composition of sulfate adenylyltransferase = [CysD]4[CysN]4
         sulfate adenylyltransferase, CysD subunit = CysD (extended summary available)
         sulfate adenylyltransferase, CysN subunit = CysN (summary available)

Summary:
Sulfate adenylyltransferase is composed of two types of subunits, CysN (53 kDa) and CysD (35 kDa) [Leyh88]. The native (390 kDa) molecular weight suggests that the enzyme is a tetramer of CysD-CysN heterodimers .

The enzyme catalyzes the activation of intracellular sulfate to adenosine 5'-phosphosulfate (APS), a reaction that generates a sulfate-phosphate anhydride linkage. This linkage facilitates an energetically-downhill entry into the subsequent metabolic fates of reduction and group transfer. The rate of APS formation is enhanced by both a protein activator and by GTP hydrolysis [Neuwald92]. The intrinsic GTPase Is believed to be the CysN subunit [Leyh92].

Gene-Reaction Schematic: ?

Credits:
Created 20-Sep-2006 by Caspi R , SRI International


Enzymatic reaction of: sulfate adenylyltransferase

Synonyms: sulfate adenylate transferase, ATP-sulfurylase, sulfurylase, ATP:sulfate adenylyltransferase, adenylylsulfate pyrophosphorylase

EC Number: 2.7.7.4

sulfate + ATP + H+ <=> adenosine 5'-phosphosulfate + diphosphate

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the Enzyme Commission system.

The reaction is favored in the direction shown.

In Pathways: superpathway of sulfate assimilation and cysteine biosynthesis , sulfate reduction I (assimilatory) , sulfate activation for sulfonation

Summary:
Studies using the hydrolysis-resistant nucleotide analogues AMPCPP and GMPPNP demonstrated that GTP hydrolysis precedes scision of the α-β bond of ATP. Product inhibition studies indicate that PPi release occurs prior to the addition of sulfate and APS formation [Liu94a].


Subunit of sulfate adenylyltransferase: sulfate adenylyltransferase, CysD subunit

Synonyms: CysD

Gene: cysD Accession Numbers: EG10186 (EcoCyc), b2752, ECK2747

Locations: cytosol

Sequence Length: 302 AAs

Molecular Weight: 35.188 kD (from nucleotide sequence)

Molecular Weight: 27 kD (experimental) [Leyh88]

GO Terms:

Biological Process: GO:0006790 - sulfur compound metabolic process Inferred from experiment [Leyh88]
GO:0006979 - response to oxidative stress Inferred from experiment [Krisko14]
GO:0000103 - sulfate assimilation Inferred by computational analysis [GOA06]
GO:0008152 - metabolic process Inferred by computational analysis [GOA01a]
GO:0019419 - sulfate reduction Inferred by computational analysis [GOA01a]
GO:0070814 - hydrogen sulfide biosynthetic process Inferred by computational analysis [UniProtGOA12]
Molecular Function: GO:0004781 - sulfate adenylyltransferase (ATP) activity Inferred from experiment Inferred by computational analysis [GOA06, GOA01, GOA01a, Leyh88]
GO:0005515 - protein binding Inferred from experiment [Leyh88]
GO:0000166 - nucleotide binding Inferred by computational analysis [UniProtGOA11a]
GO:0003824 - catalytic activity Inferred by computational analysis [GOA01a]
GO:0005524 - ATP binding Inferred by computational analysis [UniProtGOA11a]
GO:0016740 - transferase activity Inferred by computational analysis [UniProtGOA11a]
GO:0016779 - nucleotidyltransferase activity Inferred by computational analysis [UniProtGOA11a]
Cellular Component: GO:0005829 - cytosol Inferred by computational analysis [DiazMejia09]

MultiFun Terms: metabolism metabolism of other compounds sulfur metabolism

Unification Links: DIP:DIP-360N , DIP:DIP-9376N , EcoliWiki:b2752 , ModBase:P21156 , PR:PRO_000022376 , Pride:P21156 , Protein Model Portal:P21156 , RefSeq:NP_417232 , SMR:P21156 , String:511145.b2752 , Swiss-Model:P21156 , UniProt:P21156

Relationship Links: InterPro:IN-FAMILY:IPR002500 , InterPro:IN-FAMILY:IPR011784 , InterPro:IN-FAMILY:IPR014729 , Pfam:IN-FAMILY:PF01507

Summary:
CysD along with CysN are the two subunits which form sulfate adenylyltransferase [Leyh88]. This enzyme is involved in the assimilation of sulfate and catalyzes two reactions, GTP hydrolysis and activation of intracellular sulfate to adenosine 5'-phosphosulfate (APS) which generates a sulfate-phosphate anhydride linkage. This linkage facilitates an energetically-downhill entry into the subsequent metabolic fates of reduction and group transfer. The rate of APS formation is enhanced by both a protein activator and by GTP hydrolysis [Neuwald92].

Mutations in the cysD gene prevent the anabolic utilization of sulfate [Leyh88].

cysD, along with cysN and cysC, resides in the sulfate activation operon [Leyh92a].

cysD shows differential codon adaptation, resulting in differential translation efficiency signatures, in aerotolerant compared to obligate anaerobic microbes. It was therefore predicted to play a role in the oxidative stress response. A cysD deletion mutant was shown to be more sensitive than wild-type specifically to hydrogen peroxide exposure, but not other stresses [Krisko14].

Essentiality data for cysD knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enriched Yes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB Lennox Yes 37 Aerobic 7   Yes [Baba06, Comment 2]
M9 medium with 0.4% glucose No 37 Aerobic 7.2 0.27 No [Patrick07, Comment 3]
M9 medium with 1% glycerol No 37 Aerobic 7.2 0.35 No [Joyce06]
MOPS medium with 0.4% glucose Yes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]
Yes [Feist07, Comment 4]

Subunit of sulfate adenylyltransferase: sulfate adenylyltransferase, CysN subunit

Synonyms: CysN

Gene: cysN Accession Numbers: EG10194 (EcoCyc), b2751, ECK2746

Locations: cytosol

Sequence Length: 475 AAs

Molecular Weight: 52.558 kD (from nucleotide sequence)

Molecular Weight: 62 kD (experimental) [Leyh88]

pI: 5.22

GO Terms:

Biological Process: GO:0006184 - GTP catabolic process Inferred by computational analysis Inferred from experiment [Leyh92, GOA01a]
GO:0006790 - sulfur compound metabolic process Inferred from experiment Inferred by computational analysis [GOA01a, Leyh88]
GO:0000103 - sulfate assimilation Inferred by computational analysis [GOA06]
GO:0070814 - hydrogen sulfide biosynthetic process Inferred by computational analysis [UniProtGOA12]
Molecular Function: GO:0003924 - GTPase activity Inferred from experiment Inferred by computational analysis [GOA01a, Leyh92]
GO:0004781 - sulfate adenylyltransferase (ATP) activity Inferred from experiment Inferred by computational analysis [GOA06, GOA01, Leyh88]
GO:0005515 - protein binding Inferred from experiment [Leyh88]
GO:0000166 - nucleotide binding Inferred by computational analysis [UniProtGOA11a]
GO:0005524 - ATP binding Inferred by computational analysis [UniProtGOA11a]
GO:0005525 - GTP binding Inferred by computational analysis [UniProtGOA11a, GOA06, GOA01a, Leyh92]
GO:0016740 - transferase activity Inferred by computational analysis [UniProtGOA11a]
GO:0016779 - nucleotidyltransferase activity Inferred by computational analysis [UniProtGOA11a]
Cellular Component: GO:0005829 - cytosol Inferred by computational analysis [DiazMejia09]

MultiFun Terms: metabolism metabolism of other compounds sulfur metabolism

Unification Links: DIP:DIP-513N , EcoliWiki:b2751 , Mint:MINT-1247685 , ModBase:P23845 , PR:PRO_000022384 , Pride:P23845 , Protein Model Portal:P23845 , RefSeq:NP_417231 , SMR:P23845 , String:511145.b2751 , Swiss-Model:P23845 , UniProt:P23845

Relationship Links: InterPro:IN-FAMILY:IPR000795 , InterPro:IN-FAMILY:IPR004161 , InterPro:IN-FAMILY:IPR005225 , InterPro:IN-FAMILY:IPR009000 , InterPro:IN-FAMILY:IPR009001 , InterPro:IN-FAMILY:IPR011779 , InterPro:IN-FAMILY:IPR027417 , Pfam:IN-FAMILY:PF00009 , Pfam:IN-FAMILY:PF03144 , Prints:IN-FAMILY:PR00315 , Prosite:IN-FAMILY:PS00301

Summary:
CysN along with CysD are the two subunits which form sulfate adenylyltransferase [Leyh88]. CysN contains a GTP-binding consensus sequence and stimulates the synthesis of adenosine 5'-phosphosulfate (APS) by a GTPase mechanism [Leyh92]. Sulfate adenylyltransferase is involved in the assimilation of sulfate and catalyzes two reactions, GTP hydrolysis and activation of intracellular sulfate to APS which generates a sulfate-phosphate anhydride linkage. This linkage facilitates an energetically-downhill entry into the subsequent metabolic fates of reduction and group transfer. The rate of APS formation is enhanced by both a protein activator and by GTP hydrolysis [Neuwald92].

Disruption of the cysN gene prevents sulfate activation and decreases expression of the downstream cysC gene [Leyh88].

cysN, along with cysD and cysC, resides in the sulfate activation operon [Leyh92a].

Essentiality data for cysN knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enriched Yes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB Lennox Yes 37 Aerobic 7   Yes [Baba06, Comment 2]
M9 medium with 0.4% glucose No 37 Aerobic 7.2 0.27 No [Patrick07, Comment 3]
M9 medium with 1% glycerol No 37 Aerobic 7.2 0.35 No [Joyce06]
MOPS medium with 0.4% glucose Indeterminate 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]
No [Feist07, Comment 5]

References

Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554

DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114

Feist07: Feist AM, Henry CS, Reed JL, Krummenacker M, Joyce AR, Karp PD, Broadbelt LJ, Hatzimanikatis V, Palsson BO (2007). "A genome-scale metabolic reconstruction for Escherichia coli K-12 MG1655 that accounts for 1260 ORFs and thermodynamic information." Mol Syst Biol 3;121. PMID: 17593909

Gerdes03: Gerdes SY, Scholle MD, Campbell JW, Balazsi G, Ravasz E, Daugherty MD, Somera AL, Kyrpides NC, Anderson I, Gelfand MS, Bhattacharya A, Kapatral V, D'Souza M, Baev MV, Grechkin Y, Mseeh F, Fonstein MY, Overbeek R, Barabasi AL, Oltvai ZN, Osterman AL (2003). "Experimental determination and system level analysis of essential genes in Escherichia coli MG1655." J Bacteriol 185(19);5673-84. PMID: 13129938

GOA01: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

GOA01a: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

GOA06: GOA, SIB (2006). "Electronic Gene Ontology annotations created by transferring manual GO annotations between orthologous microbial proteins."

Joyce06: Joyce AR, Reed JL, White A, Edwards R, Osterman A, Baba T, Mori H, Lesely SA, Palsson BO, Agarwalla S (2006). "Experimental and computational assessment of conditionally essential genes in Escherichia coli." J Bacteriol 188(23);8259-71. PMID: 17012394

Krisko14: Krisko A, Copi T, Gabaldon T, Lehner B, Supek F (2014). "Inferring gene function from evolutionary change in signatures of translation efficiency." Genome Biol 15(3);R44. PMID: 24580753

Leyh88: Leyh TS, Taylor JC, Markham GD (1988). "The sulfate activation locus of Escherichia coli K12: cloning, genetic, and enzymatic characterization." J Biol Chem 263(5);2409-16. PMID: 2828368

Leyh92: Leyh TS, Suo Y (1992). "GTPase-mediated activation of ATP sulfurylase." J Biol Chem 1992;267(1);542-5. PMID: 1730615

Leyh92a: Leyh TS, Vogt TF, Suo Y (1992). "The DNA sequence of the sulfate activation locus from Escherichia coli K-12." J Biol Chem 1992;267(15);10405-10. PMID: 1316900

Liu94a: Liu C, Martin E, Leyh TS (1994). "GTPase activation of ATP sulfurylase: the mechanism." Biochemistry 33(8);2042-7. PMID: 8117661

Neuwald92: Neuwald AF, Krishnan BR, Brikun I, Kulakauskas S, Suziedelis K, Tomcsanyi T, Leyh TS, Berg DE (1992). "cysQ, a gene needed for cysteine synthesis in Escherichia coli K-12 only during aerobic growth." J Bacteriol 1992;174(2);415-25. PMID: 1729235

Patrick07: Patrick WM, Quandt EM, Swartzlander DB, Matsumura I (2007). "Multicopy suppression underpins metabolic evolvability." Mol Biol Evol 24(12);2716-22. PMID: 17884825

UniProtGOA11a: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

UniProtGOA12: UniProt-GOA (2012). "Gene Ontology annotation based on UniPathway vocabulary mapping."


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Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
Page generated by SRI International Pathway Tools version 18.5 on Fri Dec 19, 2014, BIOCYC13B.