Abstract
Phosphatidylinositol transfer proteins (PITPs) have been shown to play important roles in regulating a number of signal transduction pathways that couple to vesicle trafficking reactions, phosphoinositide-driven receptor-mediated signaling cascades, and development. While yeast and metazoan PITPs have been analyzed in some detail, plant PITPs remain entirely uncharacterized. We report the identification and characterization of two soybean proteins, Ssh1p and Ssh2p, whose structural genes were recovered on the basis of their abilities to rescue the viability of PITP-deficient Saccharomyces cerevisiae strains. We demonstrate that, while both Ssh1p and Ssh2p share approximately 25% primary sequence identity with yeast PITP, these proteins exhibit biochemical properties that diverge from those of the known PITPs. Ssh1p and Ssh2p represent high-affinity phosphoinositide binding proteins that are distinguished from each other both on the basis of their phospholipid binding specificities and by their substantially non-overlapping patterns of expression in the soybean plant. Finally, we show that Ssh1p is phosphorylated in response to various environmental stress conditions, including hyperosmotic stress. We suggest that Ssh1p may function as one component of a stress response pathway that serves to protect the adult plant from osmotic insult.
Full Text
The Full Text of this article is available as a PDF (518.0 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Aitken J. F., van Heusden G. P., Temkin M., Dowhan W. The gene encoding the phosphatidylinositol transfer protein is essential for cell growth. J Biol Chem. 1990 Mar 15;265(8):4711–4717. [PubMed] [Google Scholar]
- Alb J. G., Jr, Gedvilaite A., Cartee R. T., Skinner H. B., Bankaitis V. A. Mutant rat phosphatidylinositol/phosphatidylcholine transfer proteins specifically defective in phosphatidylinositol transfer: implications for the regulation of phospholipid transfer activity. Proc Natl Acad Sci U S A. 1995 Sep 12;92(19):8826–8830. doi: 10.1073/pnas.92.19.8826. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Alb J. G., Jr, Kearns M. A., Bankaitis V. A. Phospholipid metabolism and membrane dynamics. Curr Opin Cell Biol. 1996 Aug;8(4):534–541. doi: 10.1016/s0955-0674(96)80032-9. [DOI] [PubMed] [Google Scholar]
- Bankaitis V. A., Aitken J. R., Cleves A. E., Dowhan W. An essential role for a phospholipid transfer protein in yeast Golgi function. Nature. 1990 Oct 11;347(6293):561–562. doi: 10.1038/347561a0. [DOI] [PubMed] [Google Scholar]
- Bankaitis V. A., Malehorn D. E., Emr S. D., Greene R. The Saccharomyces cerevisiae SEC14 gene encodes a cytosolic factor that is required for transport of secretory proteins from the yeast Golgi complex. J Cell Biol. 1989 Apr;108(4):1271–1281. doi: 10.1083/jcb.108.4.1271. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Becker D. M., Fikes J. D., Guarente L. A cDNA encoding a human CCAAT-binding protein cloned by functional complementation in yeast. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1968–1972. doi: 10.1073/pnas.88.5.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boeke J. D., LaCroute F., Fink G. R. A positive selection for mutants lacking orotidine-5'-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet. 1984;197(2):345–346. doi: 10.1007/BF00330984. [DOI] [PubMed] [Google Scholar]
- Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
- Bowser R., Müller H., Govindan B., Novick P. Sec8p and Sec15p are components of a plasma membrane-associated 19.5S particle that may function downstream of Sec4p to control exocytosis. J Cell Biol. 1992 Sep;118(5):1041–1056. doi: 10.1083/jcb.118.5.1041. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chang J. T., Milligan S., Li Y., Chew C. E., Wiggs J., Copeland N. G., Jenkins N. A., Campochiaro P. A., Hyde D. R., Zack D. J. Mammalian homolog of Drosophila retinal degeneration B rescues the mutant fly phenotype. J Neurosci. 1997 Aug 1;17(15):5881–5890. doi: 10.1523/JNEUROSCI.17-15-05881.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chaudhary A., Gu Q. M., Thum O., Profit A. A., Qi Y., Jeyakumar L., Fleischer S., Prestwich G. D. Specific interaction of Golgi coatomer protein alpha-COP with phosphatidylinositol 3,4,5-trisphosphate. J Biol Chem. 1998 Apr 3;273(14):8344–8350. doi: 10.1074/jbc.273.14.8344. [DOI] [PubMed] [Google Scholar]
- Chinen K., Takahashi E., Nakamura Y. Isolation and mapping of a human gene (SEC14L), partially homologous to yeast SEC14, that contains a variable number of tandem repeats (VNTR) site in its 3' untranslated region. Cytogenet Cell Genet. 1996;73(3):218–223. doi: 10.1159/000134342. [DOI] [PubMed] [Google Scholar]
- Cleves A. E., McGee T. P., Whitters E. A., Champion K. M., Aitken J. R., Dowhan W., Goebl M., Bankaitis V. A. Mutations in the CDP-choline pathway for phospholipid biosynthesis bypass the requirement for an essential phospholipid transfer protein. Cell. 1991 Feb 22;64(4):789–800. doi: 10.1016/0092-8674(91)90508-v. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cleves A. E., Novick P. J., Bankaitis V. A. Mutations in the SAC1 gene suppress defects in yeast Golgi and yeast actin function. J Cell Biol. 1989 Dec;109(6 Pt 1):2939–2950. doi: 10.1083/jcb.109.6.2939. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cleves A., McGee T., Bankaitis V. Phospholipid transfer proteins: a biological debut. Trends Cell Biol. 1991 Jul;1(1):30–34. doi: 10.1016/0962-8924(91)90067-j. [DOI] [PubMed] [Google Scholar]
- Cunningham E., Tan S. K., Swigart P., Hsuan J., Bankaitis V., Cockcroft S. The yeast and mammalian isoforms of phosphatidylinositol transfer protein can all restore phospholipase C-mediated inositol lipid signaling in cytosol-depleted RBL-2H3 and HL-60 cells. Proc Natl Acad Sci U S A. 1996 Jun 25;93(13):6589–6593. doi: 10.1073/pnas.93.13.6589. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cunningham E., Thomas G. M., Ball A., Hiles I., Cockcroft S. Phosphatidylinositol transfer protein dictates the rate of inositol trisphosphate production by promoting the synthesis of PIP2. Curr Biol. 1995 Jul 1;5(7):775–783. doi: 10.1016/s0960-9822(95)00154-0. [DOI] [PubMed] [Google Scholar]
- Dickeson S. K., Lim C. N., Schuyler G. T., Dalton T. P., Helmkamp G. M., Jr, Yarbrough L. R. Isolation and sequence of cDNA clones encoding rat phosphatidylinositol transfer protein. J Biol Chem. 1989 Oct 5;264(28):16557–16564. [PubMed] [Google Scholar]
- Dormán G., Prestwich G. D. Benzophenone photophores in biochemistry. Biochemistry. 1994 May 17;33(19):5661–5673. doi: 10.1021/bi00185a001. [DOI] [PubMed] [Google Scholar]
- Dove S. K., Cooke F. T., Douglas M. R., Sayers L. G., Parker P. J., Michell R. H. Osmotic stress activates phosphatidylinositol-3,5-bisphosphate synthesis. Nature. 1997 Nov 13;390(6656):187–192. doi: 10.1038/36613. [DOI] [PubMed] [Google Scholar]
- Evan G. I., Lewis G. K., Ramsay G., Bishop J. M. Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product. Mol Cell Biol. 1985 Dec;5(12):3610–3616. doi: 10.1128/mcb.5.12.3610-3616.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fang M., Kearns B. G., Gedvilaite A., Kagiwada S., Kearns M., Fung M. K., Bankaitis V. A. Kes1p shares homology with human oxysterol binding protein and participates in a novel regulatory pathway for yeast Golgi-derived transport vesicle biogenesis. EMBO J. 1996 Dec 2;15(23):6447–6459. [PMC free article] [PubMed] [Google Scholar]
- Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
- Franzusoff A., Schekman R. Functional compartments of the yeast Golgi apparatus are defined by the sec7 mutation. EMBO J. 1989 Sep;8(9):2695–2702. doi: 10.1002/j.1460-2075.1989.tb08410.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gietz D., St Jean A., Woods R. A., Schiestl R. H. Improved method for high efficiency transformation of intact yeast cells. Nucleic Acids Res. 1992 Mar 25;20(6):1425–1425. doi: 10.1093/nar/20.6.1425. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gietz R. D., Sugino A. New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene. 1988 Dec 30;74(2):527–534. doi: 10.1016/0378-1119(88)90185-0. [DOI] [PubMed] [Google Scholar]
- Grimes H. D., Overvoorde P. J., Ripp K., Franceschi V. R., Hitz W. D. A 62-kD sucrose binding protein is expressed and localized in tissues actively engaged in sucrose transport. Plant Cell. 1992 Dec;4(12):1561–1574. doi: 10.1105/tpc.4.12.1561. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gu M., Warshawsky I., Majerus P. W. Cloning and expression of a cytosolic megakaryocyte protein-tyrosine-phosphatase with sequence homology to retinaldehyde-binding protein and yeast SEC14p. Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2980–2984. doi: 10.1073/pnas.89.7.2980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hamilton B. A., Smith D. J., Mueller K. L., Kerrebrock A. W., Bronson R. T., van Berkel V., Daly M. J., Kruglyak L., Reeve M. P., Nemhauser J. L. The vibrator mutation causes neurodegeneration via reduced expression of PITP alpha: positional complementation cloning and extragenic suppression. Neuron. 1997 May;18(5):711–722. doi: 10.1016/s0896-6273(00)80312-8. [DOI] [PubMed] [Google Scholar]
- Hammonds-Odie L. P., Jackson T. R., Profit A. A., Blader I. J., Turck C. W., Prestwich G. D., Theibert A. B. Identification and cloning of centaurin-alpha. A novel phosphatidylinositol 3,4,5-trisphosphate-binding protein from rat brain. J Biol Chem. 1996 Aug 2;271(31):18859–18868. doi: 10.1074/jbc.271.31.18859. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hay J. C., Fisette P. L., Jenkins G. H., Fukami K., Takenawa T., Anderson R. A., Martin T. F. ATP-dependent inositide phosphorylation required for Ca(2+)-activated secretion. Nature. 1995 Mar 9;374(6518):173–177. doi: 10.1038/374173a0. [DOI] [PubMed] [Google Scholar]
- Hay J. C., Martin T. F. Phosphatidylinositol transfer protein required for ATP-dependent priming of Ca(2+)-activated secretion. Nature. 1993 Dec 9;366(6455):572–575. doi: 10.1038/366572a0. [DOI] [PubMed] [Google Scholar]
- Horazdovsky B. F., Emr S. D. The VPS16 gene product associates with a sedimentable protein complex and is essential for vacuolar protein sorting in yeast. J Biol Chem. 1993 Mar 5;268(7):4953–4962. [PubMed] [Google Scholar]
- Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kader Jean-Claude. LIPID-TRANSFER PROTEINS IN PLANTS. Annu Rev Plant Physiol Plant Mol Biol. 1996 Jun;47(NaN):627–654. doi: 10.1146/annurev.arplant.47.1.627. [DOI] [PubMed] [Google Scholar]
- Kagiwada S., Kearns B. G., McGee T. P., Fang M., Hosaka K., Bankaitis V. A. The yeast BSD2-1 mutation influences both the requirement for phosphatidylinositol transfer protein function and derepression of phospholipid biosynthetic gene expression in yeast. Genetics. 1996 Jun;143(2):685–697. doi: 10.1093/genetics/143.2.685. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kearns B. G., McGee T. P., Mayinger P., Gedvilaite A., Phillips S. E., Kagiwada S., Bankaitis V. A. Essential role for diacylglycerol in protein transport from the yeast Golgi complex. Nature. 1997 May 1;387(6628):101–105. doi: 10.1038/387101a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lopez M. C., Nicaud J. M., Skinner H. B., Vergnolle C., Kader J. C., Bankaitis V. A., Gaillardin C. A phosphatidylinositol/phosphatidylcholine transfer protein is required for differentiation of the dimorphic yeast Yarrowia lipolytica from the yeast to the mycelial form. J Cell Biol. 1994 Apr;125(1):113–127. doi: 10.1083/jcb.125.1.113. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maeda T., Wurgler-Murphy S. M., Saito H. A two-component system that regulates an osmosensing MAP kinase cascade in yeast. Nature. 1994 May 19;369(6477):242–245. doi: 10.1038/369242a0. [DOI] [PubMed] [Google Scholar]
- McGee T. P., Skinner H. B., Whitters E. A., Henry S. A., Bankaitis V. A. A phosphatidylinositol transfer protein controls the phosphatidylcholine content of yeast Golgi membranes. J Cell Biol. 1994 Feb;124(3):273–287. doi: 10.1083/jcb.124.3.273. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mead D. A., Szczesna-Skorupa E., Kemper B. Single-stranded DNA 'blue' T7 promoter plasmids: a versatile tandem promoter system for cloning and protein engineering. Protein Eng. 1986 Oct-Nov;1(1):67–74. doi: 10.1093/protein/1.1.67. [DOI] [PubMed] [Google Scholar]
- Milligan S. C., Alb J. G., Jr, Elagina R. B., Bankaitis V. A., Hyde D. R. The phosphatidylinositol transfer protein domain of Drosophila retinal degeneration B protein is essential for photoreceptor cell survival and recovery from light stimulation. J Cell Biol. 1997 Oct 20;139(2):351–363. doi: 10.1083/jcb.139.2.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Murray M. G., Thompson W. F. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res. 1980 Oct 10;8(19):4321–4325. doi: 10.1093/nar/8.19.4321. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Novick P., Field C., Schekman R. Identification of 23 complementation groups required for post-translational events in the yeast secretory pathway. Cell. 1980 Aug;21(1):205–215. doi: 10.1016/0092-8674(80)90128-2. [DOI] [PubMed] [Google Scholar]
- PAULUS H., KENNEDY E. P. The enzymatic synthesis of inositol monophosphatide. J Biol Chem. 1960 May;235:1303–1311. [PubMed] [Google Scholar]
- Poon P. P., Wang X., Rotman M., Huber I., Cukierman E., Cassel D., Singer R. A., Johnston G. C. Saccharomyces cerevisiae Gcs1 is an ADP-ribosylation factor GTPase-activating protein. Proc Natl Acad Sci U S A. 1996 Sep 17;93(19):10074–10077. doi: 10.1073/pnas.93.19.10074. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Prestwich G. D., Dormán G., Elliott J. T., Marecak D. M., Chaudhary A. Benzophenone photoprobes for phosphoinositides, peptides and drugs. Photochem Photobiol. 1997 Feb;65(2):222–234. doi: 10.1111/j.1751-1097.1997.tb08548.x. [DOI] [PubMed] [Google Scholar]
- Rothstein R. J. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. doi: 10.1016/0076-6879(83)01015-0. [DOI] [PubMed] [Google Scholar]
- Salama S. R., Cleves A. E., Malehorn D. E., Whitters E. A., Bankaitis V. A. Cloning and characterization of Kluyveromyces lactis SEC14, a gene whose product stimulates Golgi secretory function in Saccharomyces cerevisiae. J Bacteriol. 1990 Aug;172(8):4510–4521. doi: 10.1128/jb.172.8.4510-4521.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sato Y., Arai H., Miyata A., Tokita S., Yamamoto K., Tanabe T., Inoue K. Primary structure of alpha-tocopherol transfer protein from rat liver. Homology with cellular retinaldehyde-binding protein. J Biol Chem. 1993 Aug 25;268(24):17705–17710. [PubMed] [Google Scholar]
- Sha B., Phillips S. E., Bankaitis V. A., Luo M. Crystal structure of the Saccharomyces cerevisiae phosphatidylinositol-transfer protein. Nature. 1998 Jan 29;391(6666):506–510. doi: 10.1038/35179. [DOI] [PubMed] [Google Scholar]
- Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Skinner H. B., Alb J. G., Jr, Whitters E. A., Helmkamp G. M., Jr, Bankaitis V. A. Phospholipid transfer activity is relevant to but not sufficient for the essential function of the yeast SEC14 gene product. EMBO J. 1993 Dec;12(12):4775–4784. doi: 10.1002/j.1460-2075.1993.tb06166.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Skinner H. B., McGee T. P., McMaster C. R., Fry M. R., Bell R. M., Bankaitis V. A. The Saccharomyces cerevisiae phosphatidylinositol-transfer protein effects a ligand-dependent inhibition of choline-phosphate cytidylyltransferase activity. Proc Natl Acad Sci U S A. 1995 Jan 3;92(1):112–116. doi: 10.1073/pnas.92.1.112. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tall E., Dormán G., Garcia P., Runnels L., Shah S., Chen J., Profit A., Gu Q. M., Chaudhary A., Prestwich G. D. Phosphoinositide binding specificity among phospholipase C isozymes as determined by photo-cross-linking to novel substrate and product analogs. Biochemistry. 1997 Jun 10;36(23):7239–7248. doi: 10.1021/bi9702288. [DOI] [PubMed] [Google Scholar]
- Tanaka S., Hosaka K. Cloning of a cDNA encoding a second phosphatidylinositol transfer protein of rat brain by complementation of the yeast sec14 mutation. J Biochem. 1994 May;115(5):981–984. doi: 10.1093/oxfordjournals.jbchem.a124448. [DOI] [PubMed] [Google Scholar]
- Thomas G. M., Cunningham E., Fensome A., Ball A., Totty N. F., Truong O., Hsuan J. J., Cockcroft S. An essential role for phosphatidylinositol transfer protein in phospholipase C-mediated inositol lipid signaling. Cell. 1993 Sep 10;74(5):919–928. doi: 10.1016/0092-8674(93)90471-2. [DOI] [PubMed] [Google Scholar]
- Vihtelic T. S., Goebl M., Milligan S., O'Tousa J. E., Hyde D. R. Localization of Drosophila retinal degeneration B, a membrane-associated phosphatidylinositol transfer protein. J Cell Biol. 1993 Sep;122(5):1013–1022. doi: 10.1083/jcb.122.5.1013. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wirtz K. W. Phospholipid transfer proteins. Annu Rev Biochem. 1991;60:73–99. doi: 10.1146/annurev.bi.60.070191.000445. [DOI] [PubMed] [Google Scholar]
