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PMID: 20389282 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S.

Crystal structure of the yeast Sac1: implications for its phosphoinositide phosphatase function.

The EMBO journal ·Vol. 29 ·No. 9 ·2010-05-05 ·Pages 1489-98

Manford A, Xia T, Saxena AK, Stefan C, Hu F, Emr SD, Mao Y

Abstract

Sac family phosphoinositide (PI) phosphatases are an essential family of CX(5)R(T/S)-based enzymes, involved in numerous aspects of cellular function such as PI homeostasis, cellular signalling, and membrane trafficking. Genetic deletions of several Sac family members result in lethality in animal models and mutations of the Sac3 gene have been found in human hereditary diseases. In this study, we report the crystal structure of a founding member of this family, the Sac phosphatase domain of yeast Sac1. The 2.0 A resolution structure shows that the Sac domain comprises of two closely packed sub-domains, a novel N-terminal sub-domain and the PI phosphatase catalytic sub-domain. The structure further shows a striking conformation of the catalytic P-loop and a large positively charged groove at the catalytic site. These findings suggest an unusual mechanism for its dephosphorylation function. Homology structural modeling of human Fig4/Sac3 allows the mapping of several disease-related mutations and provides a framework for the understanding of the molecular mechanisms of human diseases.

MeSH Terms
Amino Acid Sequence Catalytic Domain Crystallography, X-Ray Flavoproteins/chemistry,genetics Humans Models, Molecular Molecular Sequence Data Mutation Mutation, Missense Phosphoric Monoester Hydrolases/chemistry,genetics,metabolism Protein Conformation Protein Folding Saccharomyces cerevisiae/chemistry,genetics,metabolism Saccharomyces cerevisiae Proteins/chemistry,genetics,metabolism Sequence Alignment Structural Homology, Protein
Chemicals
Flavoproteins Saccharomyces cerevisiae Proteins FIG4 protein, human SAC1 protein, S cerevisiae Phosphoric Monoester Hydrolases
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Manford Andrew
Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY, USA.
Xia Tian
Saxena Ajay Kumar
Stefan Christopher
Hu Fenghua
Emr Scott D
Mao Yuxin
References (63)
63 references, click to expand
  1. The CCP4 suite: programs for protein crystallography.
    Acta Crystallogr D Biol Crystallogr. 1994 Sep 1;50(Pt 5):760-3 PMID: 15299374
  2. XtalView/Xfit--A versatile program for manipulating atomic coordinates and electron density.
    J Struct Biol. 1999 Apr-May;125(2-3):156-65 PMID: 10222271
  3. Regulation of intracellular phosphatidylinositol-4-phosphate by the Sac1 lipid phosphatase.
    Traffic. 2005 Feb;6(2):116-30 PMID: 15634212
  4. VAC14 nucleates a protein complex essential for the acute interconversion of PI3P and PI(3,5)P(2) in yeast and mouse.
    EMBO J. 2008 Dec 17;27(24):3221-34 PMID: 19037259
  5. Processing of X-ray diffraction data collected in oscillation mode.
    Methods Enzymol. 1997;276:307-26 PMID: 27754618
  6. Mutation of FIG4 causes neurodegeneration in the pale tremor mouse and patients with CMT4J.
    Nature. 2007 Jul 5;448(7149):68-72 PMID: 17572665
  7. Trimeric structure of PRL-1 phosphatase reveals an active enzyme conformation and regulation mechanisms.
    J Mol Biol. 2005 Jan 14;345(2):401-13 PMID: 15571731
  8. Web services at the European bioinformatics institute.
    Nucleic Acids Res. 2007 Jul;35(Web Server issue):W6-11 PMID: 17576686
  9. PI-loting membrane traffic.
    Nat Cell Biol. 2004 Jun;6(6):487-92 PMID: 15170460
  10. Mutations in the Saccharomyces cerevisiae gene SAC1 cause multiple drug sensitivity.
    Yeast. 1999 Aug;15(11):1111-24 PMID: 10455234
  11. Phosphoinositides in cell regulation and membrane dynamics.
    Nature. 2006 Oct 12;443(7112):651-7 PMID: 17035995
  12. Assembly of a Fab1 phosphoinositide kinase signaling complex requires the Fig4 phosphoinositide phosphatase.
    Mol Biol Cell. 2008 Oct;19(10):4273-86 PMID: 18653468
  13. Oxidation state of the active-site cysteine in protein tyrosine phosphatase 1B.
    Nature. 2003 Jun 12;423(6941):773-7 PMID: 12802339
  14. Synthesis and function of membrane phosphoinositides in budding yeast, Saccharomyces cerevisiae.
    Biochim Biophys Acta. 2007 Mar;1771(3):353-404 PMID: 17382260
  15. Solvent content of protein crystals.
    J Mol Biol. 1968 Apr 28;33(2):491-7 PMID: 5700707
  16. Interfacial binding of secreted phospholipases A(2): more than electrostatics and a major role for tryptophan.
    Curr Opin Struct Biol. 1999 Aug;9(4):428-32 PMID: 10449366
  17. The Sac1 phosphoinositide phosphatase regulates Golgi membrane morphology and mitotic spindle organization in mammals.
    Mol Biol Cell. 2008 Jul;19(7):3080-96 PMID: 18480408
  18. Crystal structure of human protein tyrosine phosphatase 1B.
    Science. 1994 Mar 11;263(5152):1397-404 PMID: 8128219
  19. An essential role for a phospholipid transfer protein in yeast Golgi function.
    Nature. 1990 Oct 11;347(6293):561-2 PMID: 2215682
  20. Crystal structure at 2.2 A resolution of the pleckstrin homology domain from human dynamin.
    Cell. 1994 Oct 21;79(2):199-209 PMID: 7954789
  21. Membrane and juxtamembrane targeting by PH and PTB domains.
    Biochim Biophys Acta. 2006 Aug;1761(8):850-67 PMID: 16807090
  22. Crystal structure of a phosphoinositide phosphatase, MTMR2: insights into myotubular myopathy and Charcot-Marie-Tooth syndrome.
    Mol Cell. 2003 Dec;12(6):1391-402 PMID: 14690594
  23. Functional studies of the mammalian Sac1 phosphoinositide phosphatase.
    Adv Enzyme Regul. 2009;49(1):75-86 PMID: 19534026
  24. Phosphoinositide 5-phosphatase Fig 4p is required for both acute rise and subsequent fall in stress-induced phosphatidylinositol 3,5-bisphosphate levels.
    Eukaryot Cell. 2006 Apr;5(4):723-31 PMID: 16607019
  25. Crystal structure of Yersinia protein tyrosine phosphatase at 2.5 A and the complex with tungstate.
    Nature. 1994 Aug 18;370(6490):571-5 PMID: 8052312
  26. Integration of Golgi trafficking and growth factor signaling by the lipid phosphatase SAC1.
    J Cell Biol. 2008 Feb 25;180(4):803-12 PMID: 18299350
  27. Kinetics of interfacial catalysis by phospholipase A2 in intravesicle scooting mode, and heterofusion of anionic and zwitterionic vesicles.
    Biochim Biophys Acta. 1986 Sep 11;860(3):435-47 PMID: 3741860
  28. The PSIPRED protein structure prediction server.
    Bioinformatics. 2000 Apr;16(4):404-5 PMID: 10869041
  29. Essential role for diacylglycerol in protein transport from the yeast Golgi complex.
    Nature. 1997 May 1;387(6628):101-5 PMID: 9139830
  30. ALSCRIPT: a tool to format multiple sequence alignments.
    Protein Eng. 1993 Jan;6(1):37-40 PMID: 8433969
  31. Mutations in the SAC1 gene suppress defects in yeast Golgi and yeast actin function.
    J Cell Biol. 1989 Dec;109(6 Pt 1):2939-50 PMID: 2687291
  32. SAC1-like domains of yeast SAC1, INP52, and INP53 and of human synaptojanin encode polyphosphoinositide phosphatases.
    J Biol Chem. 1999 May 7;274(19):12990-5 PMID: 10224048
  33. ArPIKfyve homomeric and heteromeric interactions scaffold PIKfyve and Sac3 in a complex to promote PIKfyve activity and functionality.
    J Mol Biol. 2008 Dec 26;384(4):766-79 PMID: 18950639
  34. SAC1 encodes a regulated lipid phosphoinositide phosphatase, defects in which can be suppressed by the homologous Inp52p and Inp53p phosphatases.
    J Biol Chem. 2000 Jan 14;275(2):801-8 PMID: 10625610
  35. The phosphoinositide phosphatase Sac1p controls trafficking of the yeast Chs3p chitin synthase.
    Curr Biol. 2001 Sep 18;11(18):1421-6 PMID: 11566100
  36. SAC1p is an integral membrane protein that influences the cellular requirement for phospholipid transfer protein function and inositol in yeast.
    J Cell Biol. 1993 Jul;122(1):79-94 PMID: 8314848
  37. Crystallography & NMR system: A new software suite for macromolecular structure determination.
    Acta Crystallogr D Biol Crystallogr. 1998 Sep 1;54(Pt 5):905-21 PMID: 9757107
  38. Vacuole size control: regulation of PtdIns(3,5)P2 levels by the vacuole-associated Vac14-Fig4 complex, a PtdIns(3,5)P2-specific phosphatase.
    Mol Biol Cell. 2004 Jan;15(1):24-36 PMID: 14528018
  39. Identification of a second aryl phosphate-binding site in protein-tyrosine phosphatase 1B: a paradigm for inhibitor design.
    Proc Natl Acad Sci U S A. 1997 Dec 9;94(25):13420-5 PMID: 9391040
  40. Phosphoinositide signaling and the regulation of membrane trafficking in yeast.
    Trends Biochem Sci. 2000 May;25(5):229-35 PMID: 10782093
  41. Crystal structure of the PTEN tumor suppressor: implications for its phosphoinositide phosphatase activity and membrane association.
    Cell. 1999 Oct 29;99(3):323-34 PMID: 10555148
  42. Sac1 lipid phosphatase and Stt4 phosphatidylinositol 4-kinase regulate a pool of phosphatidylinositol 4-phosphate that functions in the control of the actin cytoskeleton and vacuole morphology.
    Mol Biol Cell. 2001 Aug;12(8):2396-411 PMID: 11514624
  43. Membrane-protein interactions in cell signaling and membrane trafficking.
    Annu Rev Biophys Biomol Struct. 2005;34:119-51 PMID: 15869386
  44. The Sac1 lipid phosphatase regulates cell shape change and the JNK cascade during dorsal closure in Drosophila.
    Curr Biol. 2003 Oct 28;13(21):1882-7 PMID: 14588244
  45. Pleiotropic alterations in lipid metabolism in yeast sac1 mutants: relationship to "bypass Sec14p" and inositol auxotrophy.
    Mol Biol Cell. 1999 Jul;10(7):2235-50 PMID: 10397762
  46. Pheromone-regulated genes required for yeast mating differentiation.
    J Cell Biol. 1998 Feb 9;140(3):461-83 PMID: 9456310
  47. Redox regulation of protein tyrosine phosphatase 1B involves a sulphenyl-amide intermediate.
    Nature. 2003 Jun 12;423(6941):769-73 PMID: 12802338
  48. Dali: a network tool for protein structure comparison.
    Trends Biochem Sci. 1995 Nov;20(11):478-80 PMID: 8578593
  49. Coot: model-building tools for molecular graphics.
    Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32 PMID: 15572765
  50. The Vac14p-Fig4p complex acts independently of Vac7p and couples PI3,5P2 synthesis and turnover.
    J Cell Biol. 2006 Feb 27;172(5):693-704 PMID: 16492811
  51. Reversible oxidation of the membrane distal domain of receptor PTPalpha is mediated by a cyclic sulfenamide.
    Biochemistry. 2007 Jan 23;46(3):709-19 PMID: 17223692
  52. Modulation of sphingolipid metabolism by the phosphatidylinositol-4-phosphate phosphatase Sac1p through regulation of phosphatidylinositol in Saccharomyces cerevisiae.
    J Biol Chem. 2009 Mar 20;284(12):7588-96 PMID: 19139096
  53. Suppressors of yeast actin mutations.
    Genetics. 1989 Apr;121(4):659-74 PMID: 2656401
  54. The structure and mechanism of protein phosphatases: insights into catalysis and regulation.
    Annu Rev Biophys Biomol Struct. 1998;27:133-64 PMID: 9646865
  55. Deleterious variants of FIG4, a phosphoinositide phosphatase, in patients with ALS.
    Am J Hum Genet. 2009 Jan;84(1):85-8 PMID: 19118816
  56. Protein tyrosine phosphatases: structure-function relationships.
    FEBS J. 2008 Mar;275(5):867-82 PMID: 18298793
  57. Structure of the high affinity complex of inositol trisphosphate with a phospholipase C pleckstrin homology domain.
    Cell. 1995 Dec 15;83(6):1037-46 PMID: 8521504
  58. Functional characterization of a mammalian Sac1 and mutants exhibiting substrate-specific defects in phosphoinositide phosphatase activity.
    J Biol Chem. 2000 Nov 3;275(44):34293-305 PMID: 10887188
  59. Fab1p PtdIns(3)P 5-kinase function essential for protein sorting in the multivesicular body.
    Cell. 1998 Dec 11;95(6):847-58 PMID: 9865702
  60. Automated protein model building combined with iterative structure refinement.
    Nat Struct Biol. 1999 May;6(5):458-63 PMID: 10331874
  61. The human phosphatidylinositol phosphatase SAC1 interacts with the coatomer I complex.
    J Biol Chem. 2003 Dec 26;278(52):52689-99 PMID: 14527956
  62. SAC1 lipid phosphatase and growth control of the secretory pathway.
    Mol Biosyst. 2009 Jan;5(1):36-42 PMID: 19081929
  63. Sac phosphatase domain proteins.
    Biochem J. 2000 Sep 1;350 Pt 2:337-52 PMID: 10947947
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
1460-2075
Published
2010-05-05
Epub
2010-00-13
Pages
1489-98
Language
English
Region
England
NLM ID
8208664
PMCID
PMC2876947
Subset
IM
Grants
NIGMS NIH HHS · DMR0225180 · United States
NCRR NIH HHS · P41 RR001646 · United States
NCRR NIH HHS · RR-01646 · United States
Corrections
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