Acta Physiologiae Plantarum

, Volume 33, Issue 6, pp 2103–2116 | Cite as

Biochemical characterization of the Arabidopsis KS-type dehydrin protein, whose gene expression is constitutively abundant rather than stress dependent

  • Masakazu Hara
  • Yuri Shinoda
  • Masayuki Kubo
  • Daiju Kashima
  • Ikuo Takahashi
  • Takanari Kato
  • Tokumasa Horiike
  • Toru Kuboi
Original Paper

Abstract

Dehydrins are known as plant stress-responsive genes. Arabidopsis thaliana has 10 dehydrin genes. Among them, one of the highly expressed genes is a KS-type dehydrin (At1g54410). However, the gene product, which is a histidine-rich dehydrin whose molecular mass is 11 kDa (AtHIRD11), has not been studied. Thus, we report the biochemical characterization of the AtHIRD11 protein. Although the AtHIRD11 protein was detected in all organs of Arabidopsis, the bolting stem and the flower showed higher accumulation than the other organs, with the AtHIRD11 protein detected in the cambial zone of the stem vasculature. Most of the AtHIRD11 protein was found to be a bound form. The bound AtHIRD11 was solubilized by 1 M NaCl solution. The extracted AtHIRD11 was retained in immobilized metal-affinity chromatography, and eluted by an imidazole gradient. The native AtHIRD11 prepared from Arabidopsis was partially phosphorylated, but further phosphorylated by casein kinase 2 in vitro. Metal-binding assays indicated that Zn2+ may be the best metal for AtHIRD11 binding. These results suggest that AtHIRD11 is a metal-binding dehydrin that shows a house-keeping expression in Arabidopsis.

Keywords

Arabidopsisthaliana Dehydrin Late embryogenesis abundant proteins Metal binding 

Abbreviations

ABA

Abscisic acid

AtHIRD11

Arabidopsisthaliana histidine-rich dehydrin whose molecular mass is 11 kDa

CK2

Casein kinase 2

2D-PAGE

Two-dimensional polyacrylamide gel electrophoresis

EDTA

Ethylenediaminetetraacetic acid

EST

Expressed sequence tag

IMAC

Immobilized metal affinity chromatography

PCR

Polymerase chain reaction

RT-PCR

Reverse transcription-polymerase chain reaction

SAP

Shrimp alkaline phosphatase

SDS

Sodium dodecyl sulfate

SDS-PAGE

SDS-polyacrylamide gel electrophoresis

Supplementary material

11738_2011_749_MOESM1_ESM.pdf (77 kb)
Supplementary figures (PDF 76 kb)

References

  1. Abu-Abied M, Golomb L, Belausov E, Huang S, Geiger B, Kam Z, Staiger CJ, Sadot E (2006) Identification of plant cytoskeleton-interacting proteins by screening for actin stress fiber association in mammalian fibroblasts. Plant J 48:367–379PubMedCrossRefGoogle Scholar
  2. Alsheikh MK, Heyen BJ, Randall SK (2003) Ion binding properties of the dehydrin ERD14 are dependent upon phosphorylation. J Biol Chem 278:40882–40889PubMedCrossRefGoogle Scholar
  3. Alsheikh MK, Svensson JT, Randall SK (2005) Phosphorylation regulated ion-binding is a property shared by the acidic subclass dehydrins. Plant Cell Environ 28:1114–1122CrossRefGoogle Scholar
  4. Battaglia M, Olvera-Carrillo Y, Garciarrubio A, Campos F, Covarrubias AA (2008) The enigmatic LEA proteins and other hydrophilins. Plant Physiol 148:6–24PubMedCrossRefGoogle Scholar
  5. Bravo LA, Close TJ, Corcuera LJ, Guy CL (1999) Characterization of an 80-kDa dehydrin-like protein in barley responsive to cold acclimation. Physiol Plant 106:177–183CrossRefGoogle Scholar
  6. Brini F, Hanin M, Lumbreras V, Amara I, Khoudi H, Hassairi A, Pagès M, Masmoudi K (2007) Overexpression of wheat dehydrin DHN-5 enhances tolerance to salt and osmotic stress in Arabidopsis thaliana. Plant Cell Rep 26:2017–2026PubMedCrossRefGoogle Scholar
  7. Cheng Z, Targolli J, Huang X, Wu R (2002) Wheat LEA genes, PMA80 and PMA1959 enhance dehydration tolerance of transgenic rice (Oryza sativa L.). Mol Breed 10:71–82CrossRefGoogle Scholar
  8. Close TJ (1996) Dehydrins: emergence of a biochemical role of a family of plant dehydration proteins. Physiol Plant 97:795–803CrossRefGoogle Scholar
  9. Danyluk J, Perron A, Houde M, Limin A, Fowler B, Benhamou N, Sarhan F (1998) Accumulation of an acidic dehydrin in the vicinity of the plasma membrane during cold acclimation of wheat. Plant Cell 10:623–638PubMedCrossRefGoogle Scholar
  10. Figueras M, Pujal J, Saleh A, Save R, Pagès M, Goday A (2004) Maize Rabl7 overexpression in Arabidopsis plants promotes osmotic stress tolerance. Ann Appl Biol 144:251–257CrossRefGoogle Scholar
  11. Godoy JA, Lunar R, Torres-Schumann S, Moreno J, Rodrigo RM, Pintor-Toro JA (1994) Expression, tissue distribution and subcellular localization of dehydrin TAS14 in salt-stressed tomato plants. Plant Mol Biol 26:1921–1934PubMedCrossRefGoogle Scholar
  12. Hara M (2010) The multifunctionality of dehydrins: an overview. Plant Signal Behav 5:503–508Google Scholar
  13. Hara M, Terashima S, Fukaya T, Kuboi T (2003) Enhancement of cold tolerance and inhibition of lipid peroxidation by citrus dehydrin in transgenic tobacco. Planta 217:290–298PubMedGoogle Scholar
  14. Hara M, Fujinaga M, Kuboi T (2005) Metal binding by citrus dehydrin with histidine-rich domains. J Exp Bot 56:2695–2703PubMedCrossRefGoogle Scholar
  15. Hara M, Shinoda Y, Tanaka Y, Kuboi T (2009) DNA binding of citrus dehydrin promoted by zinc ion. Plant Cell Environ 32:532–541PubMedCrossRefGoogle Scholar
  16. Heyen BJ, Alsheikh MK, Smith EA, Torvik CF, Seals DF, Randall SK (2002) The calcium-binding activity of a vacuole-associated, dehydrin-like protein is regulated by phosphorylation. Plant Physiol 130:675–687PubMedCrossRefGoogle Scholar
  17. Hirayama T, Shinozaki K (2010) Research on plant abiotic stress responses in the post-genome era: past, present and future. Plant J 61:1041–1052PubMedCrossRefGoogle Scholar
  18. Houde M, Dallaire S, N’Dong D, Sarhan F (2004) Overexpression of the acidic dehydrin WCOR410 improves freezing tolerance in transgenic strawberry leaves. Plant Biotechnol J 2:381–387PubMedCrossRefGoogle Scholar
  19. Hundertmark M, Hincha DK (2008) LEA (Late Embryogenesis Abundant) proteins and their encoding genes in Arabidopsis thaliana. BMC Genomics 9:118PubMedCrossRefGoogle Scholar
  20. Ismail AM, Hall AE, Close TJ (1999) Allelic variation of a dehydrin gene cosegregates with chilling tolerance during seedling emergence. Proc Natl Acad Sci USA 96:13566–13570PubMedCrossRefGoogle Scholar
  21. Jiang X, Wang Y (2004) β-Elimination coupled with tandem mass spectrometry for the identification of in vivo and in vitro phosphorylation sites in maize dehydrin DHN1 protein. Biochemistry 43:15567–15576PubMedCrossRefGoogle Scholar
  22. Koag MC, Wilkens S, Fenton RD, Resnik J, Vo E, Close TJ (2009) The K-segment of maize DHN1 mediates binding to anionic phospholipid vesicles and concomitant structural changes. Plant Physiol 150:1503–1514PubMedCrossRefGoogle Scholar
  23. Kovacs D, Kalmar E, Torok Z, Tompa P (2008) Chaperone activity of ERD10 and ERD14, two disordered stress-related plant proteins. Plant Physiol 147:381–390PubMedCrossRefGoogle Scholar
  24. Krüger C, Berkowitz O, Stephan UW, Hell R (2002) A metal-binding member of the late embryogenesis abundant protein family transports iron in the phloem of Ricinus communis L. J Biol Chem 277:25062–25069PubMedCrossRefGoogle Scholar
  25. Nylander M, Svensson J, Palva ET, Welin BV (2001) Stress-induced accumulation and tissue-specific localization of dehydrins in Arabidopsis thaliana. Plant Mol Biol 45:263–279PubMedCrossRefGoogle Scholar
  26. Plana M, Itarte E, Eritja R, Goday A, Pages M, Martinez MC (1991) Phosphorylation of the maize RAB-17 protein by casein kinase 2. J Biol Chem 266:22510–22514PubMedGoogle Scholar
  27. Puhakainen T, Hess MW, Mäkelä P, Svensson J, Heino P, Palva ET (2004) Overexpression of multiple dehydrin genes enhances tolerance to freezing stress in Arabidopsis. Plant Mol Biol 54:743–753PubMedCrossRefGoogle Scholar
  28. Reyes JL, Campos F, Wei H, Arora R, Yang Y, Karlson DT, Covarrubias AA (2008) Functional dissection of hydrophilins during in vitro freeze protection. Plant Cell Environ 31:1781–1790PubMedCrossRefGoogle Scholar
  29. Röhrig H, Schmidt J, Colby T, Bräutigam A, Hufnagel P, Böhm N, Bartels D (2006) Desiccation of the resurrection plant Craterostigma plantagineum induces dynamic changes in protein phosphorylation. Plant Cell Environ 29:1606–1615PubMedCrossRefGoogle Scholar
  30. Rorat T (2006) Plant dehydrins: tissue location, structure and function. Cell Mol Biol Lett 11:536–556PubMedCrossRefGoogle Scholar
  31. Rorat T, Grygorowicz WJ, Irzykowski W, Rey P (2004) Expression of KS-type dehydrins is primarily regulated by factors related to organ type and leaf developmental stage during vegetative growth. Planta 218:878–885PubMedCrossRefGoogle Scholar
  32. Skirycz A, Inzé D (2010) More from less: plant growth under limited water. Curr Opin Biotechnol 21:197–203PubMedCrossRefGoogle Scholar
  33. Svensson J, Palva ET, Welin B (2000) Purification of recombinant Arabidopsis thaliana dehydrins by metal ion affinity chromatography. Protein Expr Purif 20:169–178PubMedCrossRefGoogle Scholar
  34. Svensson J, Ismail AM, Palva ET, Close TJ (2002) Dehydrins. In: Storey KB, Storey JM (eds) Sensing. Signaling and cell adaptation, Elsevier, pp 155–171Google Scholar
  35. Tompa P (2009) Structure and function of intrinsically disordered proteins. CRC Press, FLCrossRefGoogle Scholar
  36. Tompa P, Bánki P, Bokor M, Kamasa P, Kovács D, Lasanda G, Tompa K (2006) Protein-water and protein-buffer interactions in the aqueous solution of an intrinsically unstructured plant dehydrin: NMR intensity and DSC aspects. Biophys J 91:2243–2249PubMedCrossRefGoogle Scholar
  37. Tunnacliffe A, Wise MJ (2007) The continuing conundrum of the LEA proteins. Naturwissenschaften 94:791–812PubMedCrossRefGoogle Scholar
  38. Ueda EKM, Gout PW, Morganti L (2003) Current and prospective applications of metal ion-protein binding. J Chromatogr A 988:1–23PubMedCrossRefGoogle Scholar
  39. Wisniewski M, Webb R, Balsamo R, Close TJ, Yu XM, Griffith M (1999) Purification, immunolocalization, cryoprotective, and antifreeze activity of PCA60: a dehydrin from peach (Prunus persica). Physiol Plant 105:600–608CrossRefGoogle Scholar
  40. Yin Z, Rorat T, Szabala BM, Ziólkowska A, Malepszy S (2006) Expression of a Solanum sogarandinum SK3-type dehydrin enhances cold tolerance in transgenic cucumber seedlings. Plant Sci 170:1164–1172CrossRefGoogle Scholar

Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2011

Authors and Affiliations

  • Masakazu Hara
    • 1
  • Yuri Shinoda
    • 1
  • Masayuki Kubo
    • 1
  • Daiju Kashima
    • 1
  • Ikuo Takahashi
    • 1
  • Takanari Kato
    • 1
  • Tokumasa Horiike
    • 2
  • Toru Kuboi
    • 1
  1. 1.Faculty of AgricultureShizuoka UniversityShizuokaJapan
  2. 2.Division of Global Research LeadersShizuoka UniversityShizuokaJapan

Personalised recommendations