title: A Novel Inositol Pyrophosphate Phosphatase in Saccharomyces cerevisiae: Siw14 PROTEIN SELECTIVELY CLEAVES THE β-PHOSPHATE FROM 5-DIPHOSPHOINOSITOL PENTAKISPHOSPHATE (5PP-IP5).
abstract: Inositol pyrophosphates are high energy signaling molecules involved in cellular processes, such as energetic metabolism, telomere maintenance, stress responses, and vesicle trafficking, and can mediate protein phosphorylation. Although the inositol kinases underlying inositol pyrophosphate biosynthesis are well characterized, the phosphatases that selectively regulate their cellular pools are not fully described. The diphosphoinositol phosphate phosphohydrolase enzymes of the Nudix protein family have been demonstrated to dephosphorylate inositol pyrophosphates; however, theSaccharomyces cerevisiaehomolog Ddp1 prefers inorganic polyphosphate over inositol pyrophosphates. We identified a novel phosphatase of the recently discovered atypical dual specificity phosphatase family as a physiological inositol pyrophosphate phosphatase. Purified recombinant Siw14 hydrolyzes the β-phosphate from 5-diphosphoinositol pentakisphosphate (5PP-IP5or IP7)in vitro. In vivo,siw14Δ yeast mutants possess increased IP7levels, whereas heterologousSIW14overexpression eliminates IP7from cells. IP7levels increased proportionately whensiw14Δ was combined withddp1Δ orvip1Δ, indicating independent activity by the enzymes encoded by these genes. We conclude that Siw14 is a physiological phosphatase that modulates inositol pyrophosphate metabolism by dephosphorylating the IP7isoform 5PP-IP5to IP6.
authors.affiliation: From the Department of Biology, Georgetown University, Washington, D. C. 20057.
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Steidle
authors.affiliation: the Division of Neurosurgery, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania 19104.
authors.forename: Lucy S
authors.initials: LS
authors.lastname: Chong
authors.affiliation: the Department of Chemistry, Princeton University, Princeton, New Jersey 08544, and.
authors.forename: Mingxuan
authors.initials: M
authors.lastname: Wu
authors.affiliation: the Department of Biochemistry and Molecular and Cellular Biology, Georgetown University, Washington, D. C. 20057.
authors.forename: Elliott
authors.initials: E
authors.lastname: Crooke
authors.affiliation: the Department of Chemistry, Princeton University, Princeton, New Jersey 08544, and.
authors.forename: Dorothea
authors.initials: D
authors.lastname: Fiedler
authors.affiliation: the Division of Neurosurgery, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, resnick@email.chop.edu.
authors.forename: Adam C
authors.initials: AC
authors.lastname: Resnick
authors.affiliation: From the Department of Biology, Georgetown University, Washington, D. C. 20057, rolfesr@georgetown.edu.
authors.forename: Ronda J
authors.initials: RJ
authors.lastname: Rolfes
mesh_terms: D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D015966:Gene Expression Regulation, Fungal; D005816:Genetic Complementation Test; D007295:Inositol Phosphates / Q000378:metabolism*; D007700:Kinetics; D017856:Phosphotransferases (Phosphate Group Acceptor) / Q000235:genetics / Q000378:metabolism; D017027:Protein Tyrosine Phosphatases / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013379:Substrate Specificity
chemical_list: D007295:Inositol Phosphates; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C079849:1-diphosphoinositol pentakisphosphate; D017856:Phosphotransferases (Phosphate Group Acceptor); C099438:inositol hexakisphosphate kinase; D017027:Protein Tyrosine Phosphatases; C528878:Siw14 protein, S cerevisiae
keywords: IP7; Saccharomyces cerevisiae; Siw14; Vip1; dual-specificity phosphoprotein phosphatase; inositol phosphate; phosphatase; phosphatidylinositol; second messenger
references.citation: PLoS Genet. 2014 Sep;10(9):e1004586
references.pmid: 25254656
references.citation: Cell. 2010 Dec 10;143(6):1030-1030.e1
references.pmid: 21145466
references.citation: J Bacteriol. 1998 Apr;180(7):1841-7
references.pmid: 9537383
references.citation: Nat Protoc. 2006;1(5):2416-22
references.pmid: 17406485
references.citation: J Biol Chem. 1993 Jan 5;268(1):633-9
references.pmid: 8380170
references.citation: Nature. 2006 Mar 30;440(7084):637-43
references.pmid: 16554755
references.citation: J Biol Chem. 1994 Jul 8;269(27):17826-32
references.pmid: 8027036
references.citation: FEBS Lett. 2013 Nov 1;587(21):3464-70
references.pmid: 24021644
references.citation: Biochem J. 2009 Oct 1;423(1):109-18
references.pmid: 19614566
references.citation: Genetics. 1989 May;122(1):19-27
references.pmid: 2659436
references.citation: J Biol Chem. 2011 Sep 16;286(37):31966-74
references.pmid: 21775424
references.citation: Biochem J. 1995 Aug 15;310 ( Pt 1):279-84
references.pmid: 7646456
references.citation: J Biol Chem. 1999 Jul 30;274(31):21735-40
references.pmid: 10419486
references.citation: Chem Sci. 2013 Jan;4(1):405-410
references.pmid: 23378892
references.citation: Genetics. 2004 Feb;166(2):707-19
references.pmid: 15020461
references.citation: Science. 2011 Nov 11;334(6057):802-5
references.pmid: 22076377
references.citation: Science. 2010 May 21;328(5981):1043-6
references.pmid: 20489023
references.citation: Nature. 2002 Jan 10;415(6868):180-3
references.pmid: 11805837
references.citation: Biochem J. 2013 Jun 15;452(3):369-79
references.pmid: 23725456
references.citation: Science. 2007 Apr 6;316(5821):106-9
references.pmid: 17412958
references.citation: Science. 2009 Sep 25;325(5948):1682-6
references.pmid: 19779198
references.citation: Nature. 2006 Mar 30;440(7084):631-6
references.pmid: 16429126
references.citation: Cell Rep. 2013 May 30;3(5):1476-82
references.pmid: 23643537
references.citation: FEBS J. 2014 Jan;281(1):14-33
references.pmid: 24152294
references.citation: Mol Genet Genomics. 2011 Apr;285(4):341-54
references.pmid: 21409566
references.citation: J Mol Biol. 2007 Dec 7;374(4):899-909
references.pmid: 17976645
references.citation: Methods Mol Biol. 2012;852:97-110
references.pmid: 22328428
references.citation: J Biol Chem. 1995 Jan 27;270(4):1564-8
references.pmid: 7829485
references.citation: Angew Chem Int Ed Engl. 2014 Sep 1;53(36):9508-11
references.pmid: 25044992
references.citation: Methods Mol Biol. 2009;462:59-74
references.pmid: 19160661
references.citation: PLoS One. 2009;4(5):e5580
references.pmid: 19440344
references.citation: Proteins. 2008 Oct;73(1):241-53
references.pmid: 18433060
references.citation: J Biol Chem. 1993 Feb 25;268(6):3850-6
references.pmid: 8382679
references.citation: Angew Chem Int Ed Engl. 2014 Jul 7;53(28):7192-7
references.pmid: 24888434
references.citation: J Bacteriol. 1983 Jan;153(1):163-8
references.pmid: 6336730