Advertisement

Plant Molecular Biology

, Volume 86, Issue 4–5, pp 471–483 | Cite as

Characterization of Arabidopsis Tubby-like proteins and redundant function of AtTLP3 and AtTLP9 in plant response to ABA and osmotic stress

  • Yan Bao
  • Wei-Meng Song
  • Yan-Li Jin
  • Chun-Mei Jiang
  • Yang Yang
  • Bei Li
  • Wei-Jie Huang
  • Hua Liu
  • Hong-Xia Zhang
Article

Abstract

Tubby and Tubby-like proteins (TLPs) play essential roles in the development and function of mammal neuronal cells. In addition to the conserved carboxyl (C)-terminal Tubby domain, which is required for their plasma membrane (PM) tethering, plant TLPs also possess an amino (N)-terminal F-box domain to interact with specific Arabidopsis Skp1-like (ASK) proteins as functional SCF-type E3 ligases. Here, we report the molecular characterization of Arabidopsis TLPs (AtTLPs). β-Glucuronidase staining showed overlapped but distinct expression patterns of AtTLPs in Arabidopsis. Yeast two-hybrid assays further revealed that AtTLP1, AtTLP3, AtTLP6, AtTLP7, AtTLP9, AtTLP10 and AtTLP11 all interacted with specific ASKs, but AtTLP2, AtTLP5 and AtTLP8 did not. Subcellular localization observations in both Arabidopsis protoplasts and tobacco pollen tubes indicated that all GFP-AtTLP fusion proteins, except GFP-AtTLP8 which lacks the conserved phosphatidylinositol 4,5-bisphosphate binding sites, were targeted to the PM. Detailed studies on AtTLP3 demonstrated that AtTLP3 is a PM-tethered PIP2 binding protein which functions redundantly with AtTLP9 in abscisic acid (ABA)- and osmotic stress-mediated seed germination. Our results suggest that AtTLPs possibly work in multiple physiological and developmental processes in Arabidopsis, and AtTLP3 is also involved in ABA signaling pathway like AtTLP9 during seed germination and early seedling growth.

Keywords

Tubby Arabidopsis AtTLP ABA Stress 

Notes

Acknowledgments

We thank the Arabidopsis Biological Resource Center and the Nottingham Arabidopsis Stock Centre and GABI-Kat (Kleinboelting et al. 2012) for providing us the T-DNA insertion mutants; BiFC vectors were kindly provided by Dr. Guang Li (Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai, China). Vectors containing the ASK cDNAs were kindly provided by Dr. Nadine Schumann and Dr. Marcel Quint (Independent Junior Research Group and Department of Stress and Developmental Biology, Leibniz Institute of Plant Biochemistry, Halle, Germany). This work has been supported by the following grants: the National Natural Science Foundation of China 31000288, 31171169, 31100212, 31371228, 31370670; the National Basic Research Program of China 2010CB126600; the National Mega Project of GMO Crops 2013ZX08001003-007, 2013ZX08004002-006; and the Strategic Priority Research Program of the Chinese Academy of Sciences XDA08030108.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11103_2014_241_MOESM1_ESM.doc (17.9 mb)
Supplementary material 1 (DOC 18340 kb)
11103_2014_241_MOESM2_ESM.xls (37 kb)
Supplementary material 2 (XLS 37 kb)

References

  1. Bao Y, Wang CT, Jiang CM, Pan J, Zhang GB, Liu H, Zhang HX (2014) The tumor necrosis factor receptor-associated factor (TRAF)-like family protein SEVEN IN ABSENTIA 2 (SINA2) promotes drought tolerance in an ABA-dependent manner in Arabidopsis. New Phytol 202:174–187PubMedCrossRefGoogle Scholar
  2. Boggon TJ, Shan WS, Santagata S, Myers SC, Shapiro L (1999) Implication of tubby proteins as transcription factors by structure-based functional analysis. Science 286:2119–2125PubMedCrossRefGoogle Scholar
  3. Bu Q, Lv T, Shen H, Luong P, Wang J, Wang Z, Huang Z, Xiao L, Engineer C, Kim TH, Schroeder JI, Huq E (2014) Regulation of drought tolerance by the F-box protein MAX2 in Arabidopsis. Plant Physiol 164:424–439PubMedCrossRefPubMedCentralGoogle Scholar
  4. Cai M, Qiu DY, Yuan T, Ding XH, Li HJ, Duan L, Xu CG, Li XH, Wang SP (2008) Identification of novel pathogen-responsive cis-elements and their binding proteins in the promoter of OsWRKY13, a gene regulating rice disease resistance. Plant Cell Environ 31:86–96PubMedCrossRefGoogle Scholar
  5. Calderón-Villalobos L, Nill C, Marrocco K, Kretsch T, Schwechheimer C (2007) The evolutionarily conserved Arabidopsis thaliana F-box protein AtFBP7 is required for efficient translation during temperature stress. Gene 392:106–116PubMedCrossRefGoogle Scholar
  6. Cheng Y, Zhou W, El Sheery NI, Peters C, Li M, Wang X, Huang J (2011) Characterization of the Arabidopsis glycerophosphodiester phosphodiesterase (GDPD) family reveals a role of the plastid-localized AtGDPD1 in maintaining cellular phosphate homeostasis under phosphate starvation. Plant J 66:781–795PubMedCrossRefGoogle Scholar
  7. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743PubMedCrossRefGoogle Scholar
  8. Dreher K, Callis J (2007) Ubiquitin, hormones and biotic stress in plants. Ann Bot 99:787–822PubMedCrossRefPubMedCentralGoogle Scholar
  9. He J, Duan Y, Hua D, Fan G, Wang L, Liu Y, Chen Z, Han L, Qu LJ, Gong ZZ (2012) DEXH box RNA helicase-mediated mitochondrial reactive oxygen species production in Arabidopsis mediates crosstalk between abscisic acid and auxin signaling. Plant Cell 24:1815–1833PubMedCrossRefPubMedCentralGoogle Scholar
  10. Hua Z, Vierstra RD (2004) The cullin-RING ubiquitin-protein ligases. Annu Rev Plant Biol 62:299–334CrossRefGoogle Scholar
  11. Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusion: betaglucurodinase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907PubMedPubMedCentralGoogle Scholar
  12. Kleinboelting N, Huep G, Kloetgen A, Viehoever P, Weisshaar B (2012) GABI-Kat SimpleSearch: new features of the Arabidopsis thaliana T-DNA mutant database. Nucleic Acids Res 40:D1211–D1215PubMedCrossRefPubMedCentralGoogle Scholar
  13. Kleyn PW, Fan W, Kovats SG, Lee JJ, Pulido JC, Wu Y, Berkemeier LR, Misumi DJ, Holmgren L, Charlat O et al (1996) Identification and characterization of the mouse obesity gene tubby: a member of a novel gene family. Cell 85:281–290PubMedCrossRefGoogle Scholar
  14. Ko JH, Yang SH, Han KH (2006) Upregulation of an Arabidopsis RING-H2 gene, XERICO, confers drought tolerance through increased abscisic acid biosynthesis. Plant J 47:343–355PubMedCrossRefGoogle Scholar
  15. Koops P, Pelser S, Ignatz M, Klose C, Marrocco-Selden K, Kretsch T (2011) EDL3 is an F-box protein involved in the regulation of abscisic acid signalling in Arabidopsis thaliana. J Exp Bot 62:5547–5560PubMedCrossRefPubMedCentralGoogle Scholar
  16. Kou YJ, Qiu DY, Wang L, Li XH, Wang SP (2009) Molecular analyses of the rice tubby-like protein gene family and their response to bacterial infection. Plant Cell Rep 28:113–121PubMedCrossRefGoogle Scholar
  17. Kwak JM, Mori IC, Pei ZM, Leonhardt N, Torres MA, Dangl JL, Bloom RE, Bodde S, Jones JD, Schroeder JI (2003) NADPH oxidase AtrbohD and AtrbohF genes function in ROS-dependent ABA signaling in Arabidopsis. EMBO J 22:2623–2633PubMedCrossRefPubMedCentralGoogle Scholar
  18. Lai CP, Shaw JF (2012) Interaction analyses of Arabidopsis tubby-like proteins with ASK proteins. Bot Stud 53:447–458Google Scholar
  19. Lai CP, Lee CL, Chen PH, Wu SH, Yang CC, Shaw JF (2004) Molecular analyses of the Arabidopsis TUBBY-like protein gene family. Plant Physiol 134:1586–1597PubMedCrossRefPubMedCentralGoogle Scholar
  20. Lai CP, Chen PH, Huang JP, Tzeng YH, Chaw SM, Shaw JF (2012) Functional diversification of theTubby-like protein gene families (TULPs) during eukaryotic evolution. Biocatal Agric Biotechnol 1:2–8Google Scholar
  21. Luo DX, Bernard DG, Balk J, Hai H, Cui XF (2012) The DUF59 family gene AE7 acts in the cytosolic iron-sulfur cluster assembly pathway to maintain nuclear genome integrity in Arabidopsis. Plant Cell 24:4135–4148PubMedCrossRefPubMedCentralGoogle Scholar
  22. Moon J, Parry G, Estelle M (2004) The ubiquitin–proteasome pathway and plant development. Plant Cell 16:3181–3195PubMedCrossRefPubMedCentralGoogle Scholar
  23. Mukhopadhyay S, Jackson PK (2011) The tubby family proteins. Genome Biol 12:255CrossRefGoogle Scholar
  24. Noben-Trauth K, Naggert JK, North MA, Nishina PM (1996) A candidate gene for the mouse mutation tubby. Nature 380:534–538PubMedCrossRefGoogle Scholar
  25. Pandey GK, Cheong YH, Kim KN, Grant JJ, Li LG, Hung W, D’Angelo C, Weinl S, Kudla J, Luan S (2004) The calcium sensor calcineurin B-Like 9 modulates abscisic acid sensitivity and biosynthesis in Arabidopsis. Plant Cell 16:1912–1924PubMedCrossRefPubMedCentralGoogle Scholar
  26. Reitz MU, Bissue JK, Zocher K, Attard A, Huckelhoven R, Becker K, Imani J, Eichmann R, Schafer P (2012) The subcellular localization of tubby-like proteins and participation in stress signaling and root colonization by the mutualist Piriformospora indica. Plant Physiol 160:349–364PubMedCrossRefPubMedCentralGoogle Scholar
  27. Reitz MU, Pai S, Imani J, Schafer P (2013) New insights into the subcellular localization of tubby-like proteins and their participation in the ArabidopsisPiriformospora indica interaction. Plant Signal Behav 8:e25198PubMedCrossRefPubMedCentralGoogle Scholar
  28. Santagata S, Boggon TJ, Baird CL, Gomez CA, Zhao J, Shan WS, Myszka DG, Shapiro L (2001) G-protein signaling through tubby proteins. Science 292:2041–2050PubMedCrossRefGoogle Scholar
  29. Schumann N, Navarro-Quezada A, Ullrich K, Kuhl C, Quint M (2011) Molecular evolution and selection patterns of plant F-box proteins with C-terminal kelch repeats. Plant Physiol 155:835–850PubMedCrossRefPubMedCentralGoogle Scholar
  30. Stone SL, Callis J (2007) Ubiquitin ligases mediate growth and development by promoting protein death. Curr Opin Plant Biol 10:624–632PubMedCrossRefGoogle Scholar
  31. Wang C, Bao Y, Wang QQ, Zhang HX (2013) Introduction of the rice CYP714D1 gene into Populus inhibits expression of its homologous genes and promotes growth, biomass production and xylem fibre length in transgenic trees. J Exp Bot 64:2847–2857PubMedCrossRefPubMedCentralGoogle Scholar
  32. Yang ZF, Zhou Y, Wang XF, Gu SL, Yu JM, Liang GH, Yan CJ, Xu CW (2008) Genomewide comparative phylogenetic and molecular evolutionary analysis of tubby-like protein family in Arabidopsis, rice, and poplar. Genomics 92:246–253PubMedCrossRefGoogle Scholar
  33. Yoo SD, Cho YH, Sheen J (2007) Arabidopsis mesophyll protoplasts: a versatile cell system for transient gene expression analysis. Nature Protoco 2:1565–1572CrossRefGoogle Scholar
  34. Zhang Y, Xu W, Li Z, Deng XW, Wu W, Xue Y (2008) F-box protein DOR functions as a novel inhibitory factor for abscisic acid-induced stomatal closure under drought stress in Arabidopsis. Plant Physiol 148:2121–2133PubMedCrossRefPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Yan Bao
    • 1
  • Wei-Meng Song
    • 1
  • Yan-Li Jin
    • 1
  • Chun-Mei Jiang
    • 1
  • Yang Yang
    • 1
  • Bei Li
    • 1
  • Wei-Jie Huang
    • 1
  • Hua Liu
    • 1
  • Hong-Xia Zhang
    • 1
  1. 1.National Key Laboratory of Plant Molecular Genetics, Shanghai Institute of Plant Physiology and EcologyChinese Academy of SciencesShanghaiChina

Personalised recommendations