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Degradation of long-chain base 1-phosphate (LCBP) in Arabidopsis: functional characterization of LCBP phosphatase involved in the dehydration stress response

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Abstract

Sphingolipid metabolites, long-chain base 1-phosphates (LCBPs), are involved in ABA signaling pathways. The LCBPs synthesized by long-chain base kinase are dephosphorylated by LCBP phosphatase or degraded by LCBP lyase. Here we show that the At3g58490 gene encodes AtSPP1, a functional LCBP phosphatase. Transient expression of green fluorescent protein fusion in suspension-cultured Arabidopsis cells showed that AtSPP1 is localized in the endoplasmic reticulum. The level of dihydrosphingosine 1-phosphate was increased in loss-of-function mutants (spp1) compared with wild-type (WT) plants, suggesting a role of AtSPP1 in regulating LCBP levels. The rate of decrease in fresh weight of detached aerial parts was significantly slower in spp1 mutants than in WT plants. A stomatal closure bioassay showed that the stomata of spp1 mutants were more sensitive than the WT to ABA, suggesting that AtSPP1 is involved in guard cell signaling. However, spp1 mutants showed decreased sensitivity to ABA with respect to primary root growth but not to seed germination. The response to fumonisin B1 did not differ between the WT and spp1 mutant. A significant decrease in AtDPL1 (LCBP lyase) transcripts in spp1 mutants was observed. We conclude that AtSPP1 is a functional LCBP phosphatase that may play a role in stomatal responses through LCBP-mediated ABA signaling.

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Abbreviations

Dhs:

Dihydrosphingosine

Dhs-P:

Dihydrosphingosine 1-phosphate

FB1 :

Fumonisin B1

LCB:

Long-chain base

LCBP:

Long-chain base 1-phosphate

Phs:

Phytosphingosine

Phs-P:

Phytosphingosine 1-phosphate

Sph:

Sphingosine

Sph-P:

Sphingosine 1-phosphate

References

  • Abbas HK, Tanaka T, Duke SO, Porter JK, Wray EM, Hodges L, Sessions AE, Wang E, Merrill AH Jr, Riley RT (1994) Fumonisin-, AAL-toxin-induced disruption of sphingolipid metabolism with accumulation of free sphingoid bases. Plant Physiol 106:1085–1093

    PubMed  CAS  Google Scholar 

  • Alonso JM, Stepanova AN, Leisse TJ, Kim CJ, Chen H, Shinn P et al (2003) Genome-wide insertional mutagenesis of Arabidopsis thaliana. Science 301:653–657

    Article  PubMed  Google Scholar 

  • Baudin A, Ozier-Kalogeropoulos O, Denouel A, Lacroute F, Cullin C (1993) A simple efficient method for direct gene deletion in Saccharomyces cerevisiae. Nucleic Acids Res 21:3329–3330

    Article  PubMed  CAS  Google Scholar 

  • Brandwagt BF, Kneeppers TJ, Nijkamp HJ, Hillie J (2002) Overexpression of the tomato Asc-1 gene mediates high insensitivity to AAL toxins fumonisin B1 in tomato hairy roots confers resistance to Alternaria alternata fsp lycopersici in Nicotiana umbratica plants. Mol Plant Microb Interact 15:35–42

    Article  CAS  Google Scholar 

  • Brindley DN, Waggoner DW (1998) Mammalian lipid phosphate phosphohydrolases. J Biol Chem 273:24281–24284

    Article  PubMed  CAS  Google Scholar 

  • Chen M, Markham JE, Li J, Dietrich CR, Jaworksi JG, Cahoon EB (2008) Sphingolipid long-chain base hydroxylation is important for growth regulation of sphingolipid content composition in Arabidopsis. Plant Cell 20:1862–1878

    Article  PubMed  CAS  Google Scholar 

  • Chiu W, Niwa Y, Zeng W, Hirano T, Kobayashi H, Sheen J (1996) Engineered GFP as a vital reporter in plants. Curr Biol 6:325–330

    Article  PubMed  CAS  Google Scholar 

  • Coursol S, Fan LM, Le Stunff H, Spiegel S, Gilroy S, Assmann SM (2003) Sphingolipid signalling in Arabidopsis guard cells involves heterotrimeric G proteins. Nature 423:651–654

    Article  PubMed  CAS  Google Scholar 

  • Coursol S, Le Stunff H, Lynch DV, Gilroy S, Assmann SM, Spiegel S (2005) Arabidopsis sphingosine kinase the effects of phytosphingosine-1-phosphate on stomatal aperture. Plant Physiol 137:724–737

    Article  PubMed  CAS  Google Scholar 

  • Crowther GJ, Lynch DV (1997) Characterization of sphinganine kinase activity in corn shoot microsomes. Arch Biochem Biophys 337:284–290

    Article  PubMed  CAS  Google Scholar 

  • Cutler S, Ghassemian M, Bonetta D, Cooney S, McCourt P (1996) A protein farnesyl transferase involved in abscisic acid signal transduction in Arabidopsis. Science 273:1239–1241

    Article  PubMed  CAS  Google Scholar 

  • Gietz RD, Schiestl RH, Willems AR, Woods RA (1995) Studies on the transformation of intact yeast cells by the LiAc/SS-DNA/PEG procedure. Yeast 11:355–360

    Article  PubMed  CAS  Google Scholar 

  • Gosti F, Beaudoin N, Serizet C, Webb AA, Vartanian N, Giraudat J (1999) ABI1 protein phosphatase 2C is a negative regulator of abscisic acid signaling. Plant Cell 11:1897–1910

    Article  PubMed  CAS  Google Scholar 

  • Imai H, Nishiura H (2005) Phosphorylation of sphingoid long-chain bases in Arabidopsis: functional characterization expression of the first sphingoid long-chain base kinase gene in plants. Plant Cell Physiol 46:375–380

    Article  PubMed  CAS  Google Scholar 

  • Kihara A, Sakuraba H, Ikeda M, Denpoh A, Igarashi Y (2008) Membrane topology essential amino acid residues of Phs1, a 3-hydroxyacyl-CoA dehydratase involved in very long-chain fatty acid elongation. J Biol Chem 283:11199–11209

    Article  PubMed  CAS  Google Scholar 

  • Kinoshita T, Doi M, Suetsugu N, Kagawa T, Wada M, Shimazaki K (2001) Phot1, phot2 mediate blue light regulation of stomatal opening. Nature 414:656–660

    Article  PubMed  CAS  Google Scholar 

  • Kuroyanagi M, Yamada K, Hatsugai N, Kondo M, Nishimura M, Hara-Nishimura I (2005) Vacuolar processing enzyme is essential for mycotoxin-induced cell death in Arabidopsis thaliana. J Biol Chem 280:32914–32920

    Article  PubMed  CAS  Google Scholar 

  • Le Stunff H, Peterson C, Thornton R, Milstien S, Mandala SM, Spiegel S (2002) Characterization of murine sphingosine-1-phosphate phosphohydrolase. J Biol Chem 277:8920–8927

    Article  PubMed  Google Scholar 

  • Mandala SM, Thornton R, Tu Z, Kurtz MB, Nickels J, Broach J, Menzeleev R, Spiegel S (1998) Sphingoid base 1-phosphate phosphatase: a key regulator of sphingolipid metabolism stress response. Proc Natl Acad Sci USA 95:150–155

    Article  PubMed  CAS  Google Scholar 

  • Mandala SM, Thornton R, Galve-Roperh I, Poulton S, Peterson C, Olivera A, Bergstrom J, Kurtz MB, Spiegel S (2000) Molecular cloning characterization of a lipid phosphohydrolase that degrades sphingosine-1-phosphate induces cell death. Proc Natl Acad Sci USA 97:7859–7864

    Article  PubMed  CAS  Google Scholar 

  • Mao C, Wadleigh M, Jenkins GM, Hannun YA, Obeid LM (1997) Identification characterization of Saccharomyces cerevisiae dihydrosphingosine-1-phosphate phosphatase. J Biol Chem 272:28690–28694

    Article  PubMed  CAS  Google Scholar 

  • Mao C, Saba JD, Obeid LM (1999) The dihydrosphingosine-1-phosphate phosphatases of Saccharomyces cerevisiae are important regulators of cell proliferation heat stress responses. Biochem J 342:667–675

    Article  PubMed  CAS  Google Scholar 

  • Marion J, Bach L, Bellec Y, Meyer C, Gissot L, Faure JD (2008) Systematic analysis of protein subcellular localization interaction using high-throughput transient transformation of Arabidopsis seedlings. Plant J 56:169–179

    Article  PubMed  CAS  Google Scholar 

  • Markham JE, Jaworski JG (2007) Rapid measurement of sphingolipids from Arabidopsis thaliana by reversed-phase high-performance liquid chromatography coupled to electrospray ionization tandem mass spectrometry. Rapid Commun Mass Spectrom 21:1304–1314

    Article  PubMed  CAS  Google Scholar 

  • Nakamura Y, Tsuchiya M, Ohta H (2007) Plastidic phosphatidic acid phosphatases identified in a distinct subfamily of lipid phosphate phosphatases with prokaryotic origin. J Biol Chem 282:29013–29021

    Article  PubMed  CAS  Google Scholar 

  • Ng CK, Carr K, McAinsh MR, Powell B, Hetherington AM (2001) Drought-induced guard cell signal transduction involves sphingosine-1-phosphate. Nature 410:596–599

    Article  PubMed  CAS  Google Scholar 

  • Nishikawa M, Hosokawa K, Ishiguro M, Minamioka H, Tamura K, Hara-Nishimura I, Takahashi Y, Shimazaki K, Imai H (2008) Degradation of sphingoid long-chain base 1-phosphates (LCB-1Ps): functional characterization expression of AtDPL1 encoding LCB-1P lyase involved in the dehydration stress response in Arabidopsis. Plant Cell Physiol 49:1758–1763

    Article  PubMed  CAS  Google Scholar 

  • Ogawa C, Kihara A, Gokoh M, Igarashi Y (2003) Identification characterization of a novel human sphingosine-1-phosphate phosphohydrolase hSPP2. J Biol Chem 278:1268–1272

    Article  PubMed  CAS  Google Scholar 

  • Pierrugues O, Brutesco C, Oshiro J, Gouy M, Deveaux Y, Carman GM, Thuriaux P, Kazmaier M (2001) Lipid phosphate phosphatases in Arabidopsis. Regulation of the AtLPP1 gene in response to stress. J Biol Chem 276:20300–20308

    Article  PubMed  CAS  Google Scholar 

  • Robinson JS, Klionsky DJ, Banta LM, Emr SD (1988) Protein sorting in Saccharomyces cerevisiae: isolation of mutants defective in the delivery processing of multiple vacuolar hydrolases. Mol Cell Biol 8:4936–4948

    PubMed  CAS  Google Scholar 

  • Saba JD, Nara F, Bielawska A, Garrett S, Hunnun YA (1997) The BST1 gene of Saccharomyces cerevisiae is the sphingosine-1-phosphate lyase. J Biol Chem 272:26087–26090

    Article  PubMed  CAS  Google Scholar 

  • Shi L, Bielawski J, Mu J, Dong H, Teng C, Zhang J, Yang X, Tomishige N, Hanada K, Hannun YA, Zuo J (2007) Involvement of sphingoid bases in mediating reactive oxygen intermediate production programmed cell death in Arabidopsis. Cell Res 17:1–11

    Article  Google Scholar 

  • Spassieva SD, Markham JE, Hille J (2002) The plant disease resistance gene Asc-1 prevents disruption of sphingolipid metabolism during AAL-toxin-induced programmed cell death. Plant J 32:561–572

    Article  PubMed  CAS  Google Scholar 

  • Tsegaye Y, Richardson CG, Bravo JE, Mulcahy BJ, Lynch DV, Markham JE, Jaworski JG, Chen M, Cahoon EB, Dunn TM (2007) Arabidopsis mutants lacking long chain base phosphate lyase are fumonisin-sensitive accumulate trihydroxy-18:1 long chain base phosphate. J Biol Chem 282:28195–28206

    Article  PubMed  CAS  Google Scholar 

  • Ueda T, Yamaguchi M, Uchimiya H, Nakano A (2001) Ara6, a plant-unique novel type Rab GTPase, functions in the endocytic pathway of Arabidopsis thaliana. EMBO J 20:4730–4741

    Google Scholar 

  • Wang H, Li J, Bostock RM, Gilchrist DG (1996) Apoptosis: a functional paradigm for programmed plant cell death induced by a host selective phytotoxin invoked during development. Plant Cell 8:375–391

    Article  PubMed  CAS  Google Scholar 

  • Worrall D, Liang Y-K, Alvarez S, Holroyd GH, Spiegel S, Panagopulos M, Gray JE, Hetherington AM (2008) Involvement of sphingosine kinase in plant cell signaling. Plant J 56:64–72

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

The authors thank Mr. Sentaro Tanaka, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Japan, for his technical assistance. RAFL clones were obtained from RIKEN Genomic Sciences Center (Yokohama, Kanagawa, Japan). We thank Arabidopsis Biological Resource Center (Ohio State University, Columbus, OH, USA) for providing spp1-1 and spp1-2 seeds, and Dr. Yasuo Niwa (University of Shizuoka, Shizuoka, Japan) for the original sGFP(S65T) vector. We also thank Chihiro Iwami, Naoko Shimada, Masayuki Watanabe, Akiko Iwamoto and Mai Ishiguro for their contributions to our study.

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Correspondence to Hiroyuki Imai.

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Nakagawa, N., Kato, M., Takahashi, Y. et al. Degradation of long-chain base 1-phosphate (LCBP) in Arabidopsis: functional characterization of LCBP phosphatase involved in the dehydration stress response. J Plant Res 125, 439–449 (2012). https://doi.org/10.1007/s10265-011-0451-9

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