Regulation of Yeast Ectoapyrase Ynd1p Activity by Activator Subunit Vma13p of Vacuolar H+-ATPase*

CD39-like ectoapyrases are involved in protein and lipid glycosylation in the Golgi lumen of Saccharomyces cerevisiae. By using a two-hybrid screen, we found that an activator subunit (Vma13p) of yeast vacuolar H+-ATPase (V-ATPase) binds to the cytoplasmic domain of Ynd1p, a yeast ectoapyrase. Interaction of Ynd1p with Vma13p was demonstrated by direct binding and co-immunoprecipitation. Surprisingly, the membrane-bound ADPase activity of Ynd1p in a vma13Δmutant was drastically increased compared with that of Ynd1p inVMA13 cells. A similar increase in the apyrase activity of Ynd1p was found in a vma1Δ mutant, in which the catalytic subunit A of V-ATPase is missing, and the membrane peripheral subunits including Vma13p are dissociated from the membranes. However, the E286Q mutant of VMA1, which assembles inactive V-ATPase complex including Vma13p in the membrane, retained wild type levels of Ynd1p activity, demonstrating that the presence of Vma13p rather than the function of V-ATPase in the membrane represses Ynd1p activity. These results suggest that association of Vma13p with the cytoplasmic domain of Ynd1p regulates its apyrase activity in the Golgi lumen.

[1]  G. Guidotti,et al.  A Yeast Golgi E-type ATPase with an Unusual Membrane Topology* , 1999, The Journal of Biological Chemistry.

[2]  Patricia D. Christie,et al.  Targeted disruption of cd39/ATP diphosphohydrolase results in disordered hemostasis and thromboregulation , 1999, Nature Medicine.

[3]  H. Elsässer,et al.  A human intracellular apyrase-like protein, LALP70, localizes to lysosomal/autophagic vacuoles. , 1999, Journal of cell science.

[4]  Y. Jigami,et al.  YND1, a Homologue of GDA1, Encodes Membrane-bound Apyrase Required for Golgi N- andO-Glycosylation in Saccharomyces cerevisiae * , 1999, Journal of Biological Chemistry.

[5]  T. Rapoport,et al.  BiP Acts as a Molecular Ratchet during Posttranslational Transport of Prepro-α Factor across the ER Membrane , 1999, Cell.

[6]  N. Nelson,et al.  Vacuolar and plasma membrane proton-adenosinetriphosphatases. , 1999, Physiological reviews.

[7]  M. Forgac.  Structure and Properties of the Clathrin-Coated Vesicle and Yeast Vacuolar V-ATPases , 1999, Journal of bioenergetics and biomembranes.

[8]  P. Kane.  Biosynthesis and Regulation of the Yeast Vacuolar H+-ATPase , 1999, Journal of bioenergetics and biomembranes.

[9]  A. Johnson,et al.  The translocon: a dynamic gateway at the ER membrane. , 1999, Annual review of cell and developmental biology.

[10]  M. Forgac.  Structure, function and regulation of the vacuolar (H+)‐ATPases , 1998, FEBS letters.

[11]  K. Titani,et al.  Placental ecto-ATP diphosphohydrolase: its structural feature distinct from CD39, localization and inhibition on shear-induced platelet aggregation. , 1998, International journal of hematology.

[12]  G. Guidotti,et al.  The Transmembrane Domains of Ectoapyrase (CD39) Affect Its Enzymatic Activity and Quaternary Structure* , 1998, The Journal of Biological Chemistry.

[13]  Thomas M. Smith,et al.  Cloning, sequencing, and expression of a human brain ecto-apyrase related to both the ecto-ATPases and CD39 ecto-apyrases , 1998 .

[14]  A. Knowles,et al.  Molecular Cloning of the Chicken Oviduct Ecto-ATP-Diphosphohydrolase* , 1998, The Journal of Biological Chemistry.

[15]  A. Frischauf,et al.  The CD39-like gene family: identification of three new human members (CD39L2, CD39L3, and CD39L4), their murine homologues, and a member of the gene family from Drosophila melanogaster. , 1998, Genomics.

[16]  F. Brodsky,et al.  Interactions between HIV1 Nef and vacuolar ATPase facilitate the internalization of CD4. , 1998, Immunity.

[17]  G. Guidotti,et al.  Golgi Localization and Functional Expression of Human Uridine Diphosphatase* , 1998, The Journal of Biological Chemistry.

[18]  P. Tai,et al.  When an ATPase Is Not an ATPase: at Low Temperatures the C-Terminal Domain of the ABC Transporter CvaB Is a GTPase , 1998, Journal of bacteriology.

[19]  P. Robbins,et al.  Transporters of nucleotide sugars, ATP, and nucleotide sulfate in the endoplasmic reticulum and Golgi apparatus. , 1998, Annual review of biochemistry.

[20]  G. Guidotti,et al.  Characterization of brain ecto-apyrase: evidence for only one ecto-apyrase (CD39) gene. , 1997, Brain research. Molecular brain research.

[21]  C. Abeijon,et al.  Transporters of nucleotide sugars, nucleotide sulfate and ATP in the Golgi apparatus. , 1997, Trends in biochemical sciences.

[22]  P. Kane,et al.  Site-directed Mutagenesis of the Yeast V-ATPase A Subunit* , 1997, The Journal of Biological Chemistry.

[23]  C. Kennedy,et al.  Neuronal release of soluble nucleotidases and their role in neurotransmitter inactivation , 1997, nature.

[24]  A. Marcus,et al.  The endothelial cell ecto-ADPase responsible for inhibition of platelet function is CD39. , 1997, The Journal of clinical investigation.

[25]  T. Kirley.  Complementary DNA Cloning and Sequencing of the Chicken Muscle Ecto-ATPase , 1997, The Journal of Biological Chemistry.

[26]  T. Stevens,et al.  Structure, function and regulation of the vacuolar (H+)-ATPase. , 1997, Annual review of cell and developmental biology.

[27]  F. Bach,et al.  Identification and Characterization of CD39/Vascular ATP Diphosphohydrolase* , 1996, The Journal of Biological Chemistry.

[28]  E. Craig,et al.  Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast. , 1996, Genetics.

[29]  Ting-Fang Wang,et al.  CD39 Is an Ecto-(Ca,Mg)-apyrase (*) , 1996, The Journal of Biological Chemistry.

[30]  T. Graham,et al.  Sorting of yeast alpha 1,3 mannosyltransferase is mediated by a lumenal domain interaction, and a transmembrane domain signal that can confer clathrin-dependent Golgi localization to a secreted protein. , 1995, Molecular biology of the cell.

[31]  L. Plesner.  Ecto-ATPases: identities and functions. , 1995, International review of cytology.

[32]  H. Zimmermann.  Signalling via ATP in the nervous system , 1994, Trends in Neurosciences.

[33]  Y. Anraku,et al.  Chemiosmotic coupling of ion transport in the yeast vacuole: Its role in acidification inside organelles , 1994, Journal of bioenergetics and biomembranes.

[34]  P. Philippsen,et al.  New heterologous modules for classical or PCR‐based gene disruptions in Saccharomyces cerevisiae , 1994, Yeast.

[35]  S. Emr,et al.  Clathrin-dependent localization of alpha 1,3 mannosyltransferase to the Golgi complex of Saccharomyces cerevisiae , 1994, The Journal of cell biology.

[36]  B. Taillon,et al.  STV1 gene encodes functional homologue of 95-kDa yeast vacuolar H(+)-ATPase subunit Vph1p. , 1994, The Journal of biological chemistry.

[37]  J. Miret,et al.  The Golgi guanosine diphosphatase is required for transport of GDP-mannose into the lumen of Saccharomyces cerevisiae Golgi vesicles. , 1994, The Journal of biological chemistry.

[38]  T. Stevens,et al.  VMA13 encodes a 54-kDa vacuolar H(+)-ATPase subunit required for activity but not assembly of the enzyme complex in Saccharomyces cerevisiae. , 1993, The Journal of biological chemistry.

[39]  P. Robbins,et al.  Guanosine diphosphatase is required for protein and sphingolipid glycosylation in the Golgi lumen of Saccharomyces cerevisiae , 1993, The Journal of cell biology.

[40]  I. Mellman.  The importance of being acid: the role of acidification in intracellular membrane traffic. , 1992, The Journal of experimental biology.

[41]  C. J. Roberts,et al.  Membrane protein sorting in the yeast secretory pathway: evidence that the vacuole may be the default compartment , 1992, The Journal of cell biology.

[42]  P. Kane,et al.  Subunit composition, biosynthesis, and assembly of the yeast vacuolar proton-translocating ATPase , 1992, Journal of bioenergetics and biomembranes.

[43]  H Zimmermann,et al.  5'-Nucleotidase: molecular structure and functional aspects. , 1992, The Biochemical journal.

[44]  D. Botstein,et al.  The VPH1 gene encodes a 95-kDa integral membrane polypeptide required for in vivo assembly and activity of the yeast vacuolar H(+)-ATPase. , 1992, The Journal of biological chemistry.

[45]  P. Kane,et al.  Assembly and targeting of peripheral and integral membrane subunits of the yeast vacuolar H(+)-ATPase. , 1992, The Journal of biological chemistry.

[46]  Jeffrey H. Miller,et al.  A short course in bacterial genetics , 1992 .

[47]  S. Emr,et al.  A novel protein kinase homolog essential for protein sorting to the yeast lysosome-like vacuole , 1991, Cell.

[48]  P. Kane,et al.  Protein splicing converts the yeast TFP1 gene product to the 69-kD subunit of the vacuolar H(+)-adenosine triphosphatase. , 1990, Science.

[49]  P. Kane,et al.  Protein targeting to the yeast vacuole. , 1989, Trends in biochemical sciences.

[50]  S. Fields,et al.  A novel genetic system to detect protein–protein interactions , 1989, Nature.

[51]  M. Forgac.  Structure and function of vacuolar class of ATP-driven proton pumps. , 1989, Physiological reviews.

[52]  T. Stevens,et al.  Characterization of genes required for protein sorting and vacuolar function in the yeast Saccharomyces cerevisiae. , 1989, The EMBO journal.

[53]  R. D. Gietz,et al.  New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. , 1988, Gene.

[54]  A. McDowall,et al.  Nucleolar and nuclear envelope proteins of the yeast Saccharomyces cerevisiae. , 1988, European journal of cell biology.

[55]  J. Thorner,et al.  Isolation of the putative structural gene for the lysine-arginine-cleaving endopeptidase required for processing of yeast prepro-α-factor , 1984, Cell.

[56]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[57]  E. Brandan,et al.  Orientation and role of nucleosidediphosphatase and 5'-nucleotidase in Golgi vesicles from rat liver. , 1982, Biochemistry.

[58]  G. Fink,et al.  Methods in yeast genetics , 1979 .

[59]  A. White,et al.  The role of nucleoside diphosphatase in a uridine nucleotide cycle associated with lactose synthesis in rat mammary-gland Golgi apparatus. , 1977, The Biochemical journal.

[60]  D. G. Herries,et al.  Some kinetic properties of human-milk galactosyl transferase. , 1974, European journal of biochemistry.