In Silico and Deletion Analysis of Upstream Promoter Fragment of S-Adenosyl Homocysteine Hydrolase (SAHH1) Gene of Arabidopsis Leads to the Identification of a Fragment Capable of Driving Gene Expression in Developing Seeds and Anthers

  • T. P. Sujatha
  • C. Sivanandan
  • S. R. Bhat
  • R. Srinivasan
Article
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Abstract

S-adenosyl homocysteine hydrolase (SAHH) is a key enzyme in methylation metabolism of eukaryotes. A 1585 by fragment upstream to ATG of SAHH1 gene, was fused with a promoter-less β-Glucuronidase (GUS) gene and mobilized into Arabidopsis by Agrobacterium-mediated floral transformation to generate transgenic Arabidopsis. This fragment was found to drive constitutive expression of GUS in T2 progeny of transgenic Arabidopsis. In silico analysis of the promoter region of SAHH1 suggested the presence of several cis-regulatory motifs including seed-specific motifs as well as anther-specific motifs in the 376 by (upstream to TSS of SAHH1) promoter fragment. Based on the partial deletion analysis carried out in the promoter region of SAHH1 (At4gl3940) this 376 by promoter fragment was found to be capable of driving GUS expression in developing seeds and in some anthers/micros pores.

Key words

SAHH1 promoter methylation GUS seed-specific 

Abbreviations

ABA

Abscisic acid

ADK

Adenosine kinase

HOG 1

Homology-dependent gene silencing1

GUS

β-Glucuronidase

RNAi

RNA interference

SAHH-S

adenosyl homocysteine hydrolase

SAM-S

adenosyl methionine

TSS

transcription start site

UTR

untranslated region

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References

  1. 1.
    Stepkowski T, Brzeziński K, Legocki AB, Jaskólski M & Béna G, Mol Phylogenet Evol, 34 (2005) 15.PubMedCrossRefGoogle Scholar
  2. 2.
    Mull L, Ebbs ML & Bender JA, Genetics, 174 (2006) 1161.PubMedCrossRefGoogle Scholar
  3. 3.
    Kawalleck P, Plesch G, Hahlbrock K, & Somssich IE, Proc Natl Acad Sci, USA, 89 (1992) 4713.PubMedCrossRefGoogle Scholar
  4. 4.
    Tanaka H, Masuta C, Kataoka J, Kuwata S, Koiwai A & Noma M, Plant Sci, 113 (1996) 167.CrossRefGoogle Scholar
  5. 5.
    Weretilnyk EA, Alexander KJ, Drebenstedt M, Snider JD, Summers PS & Moffatt BA, Plant Physiol, 125 (2001) 856.PubMedCrossRefGoogle Scholar
  6. 6.
    Zheng J, Zhao J, Tao Y & Fu J, Plant Mol Biol, 55 (2004) 807.PubMedGoogle Scholar
  7. 7.
    Abrahams S, Hayes CM & Watson JIM, Plant Mol Biol, 27 (1995) 513.PubMedCrossRefGoogle Scholar
  8. 8.
    Medina-Suarez R, Manning K, Fletcher J, Aked J, Bird CR & Seymour GB, Plant Physiol, 115 (1997) 453.PubMedCrossRefGoogle Scholar
  9. 9.
    Hadfield KA, Dang T, Guis M, Pech J-C, Bouzayen M & Bennett AB, Plant Physiol, 122 (2000) 977.PubMedCrossRefGoogle Scholar
  10. 10.
    Radchuk VV, Sreenivasulu N, Radchuk RI, Wobus U & Weschke W, Plant Mol Biol, 59 (2005) 289.PubMedCrossRefGoogle Scholar
  11. 11.
    Rocha PSCF, Sheikh M, Melchiorre R, Fagard M, Bouter S, Loach R, Moffatt B, Wagner C, Vaucheret H & Furner I, Plant Cell, 17 (2005) 404.PubMedCrossRefGoogle Scholar
  12. 12.
    Li C-H, Jiang S-M, Wang L-J & Chen X-Y, TAIR accession Publication:501721780, 2007.Google Scholar
  13. 13.
    Sivanandan C, Ph.D. thesis, Indian Agricultural Research Institute, (2004).Google Scholar
  14. 14.
    Sivanandan C, Sujatha TP, Anand Mohan Prasad, Resminath R, Thakare, DR, Bhat SR & Srinivasan, Biochim Biophys Acta, 1731 (2005) 202.PubMedCrossRefGoogle Scholar
  15. 15.
    Sambrook J & Russell DW, Molecular cloning: A laboratory manual (3-Volume Set), Cold Spring Harbor Press, USA, (2001).Google Scholar
  16. 16.
    Murashige T & Skoog F, Physiol Plant, 15 (1962) 473.CrossRefGoogle Scholar
  17. 17.
    Clough S & Bent A, Plant J, 16 (1998) 735.PubMedCrossRefGoogle Scholar
  18. 18.
    Jefferson RA, Kavanagh TA & Bevan MW, EMBO J, 6 (1987) 3901.PubMedGoogle Scholar
  19. 19.
    Higo K, Ugawa Y, Iwamoto M & Korenaga T, Nucleic Acids Res, 27 (1999) 297.PubMedCrossRefGoogle Scholar
  20. 20.
    Cheung TH, Kwan YL, Hamady M & Liu X, Genome Biol, 7 (2006) R97.PubMedCrossRefGoogle Scholar
  21. 21.
    Shen B. Carneiro N, Torres-Jerez I, Stevenson R, McCreery T, Helentjaris T, Baysdorfer C, Almira E, Ferl R, Habben J & Larkins B, Plant Mol Biol, 26 (1994) 1085.PubMedCrossRefGoogle Scholar
  22. 22.
    Pereira LAR, Todorova M, Cai X,. Makaroff CA, Emery RJN & Moffatt BA, J Exp Bot, 58 (2007) 1083.PubMedCrossRefGoogle Scholar
  23. 23.
    Allen RD, Bernier F, Lessard, PA & Beachy RN, Plant Cell, 1 (1989) 623.PubMedGoogle Scholar
  24. 24.
    Thomas TL, Plant Cell, 5 (1993) 1401.PubMedGoogle Scholar
  25. 25.
    Washida H, Wu CY, Suzuki A, Yamonouchi U, Akihama T, Harada K & Takaiwa F, Plant Mol Biol, 40 (1999) 1.PubMedCrossRefGoogle Scholar
  26. 26.
    Wasserman WW, Palumbo M, Thompson W, Fickett JW & Lawrence CE, Nat Genet, 26 (2000) 225.PubMedCrossRefGoogle Scholar
  27. 27.
    Hill TA, Day CD, Zondlo SC, Thackeray AG, Irish VF, Development, 125 (1998) 1711.PubMedGoogle Scholar
  28. 28.
    Koch MA, Weisshaar B, Kroymann J, Haubold B & Mitchell-Olds T, Mol Biol Evol, 18 (2001) 1882.PubMedCrossRefGoogle Scholar
  29. 29.
    Weretilnyk EA, Alexander KJ, Drebenstedt M, Snider JD, Summers PS & Moffatt BA, Plant Physiol, 125 (2001) 856.PubMedCrossRefGoogle Scholar
  30. 30.
    Zheng J, Zhao J, Tao Y & Fu J, Plant Mol Biol, 55 (2004) 807.PubMedGoogle Scholar
  31. 31.
    Arabidopsis Genome Initiative, Nature, 408 (2000) 796.CrossRefGoogle Scholar
  32. 32.
    Ohno S, Evolution by gene duplication, Springer-Verlag, New York, (1970).Google Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • T. P. Sujatha
    • 1
  • C. Sivanandan
    • 1
  • S. R. Bhat
    • 1
  • R. Srinivasan
    • 1
  1. 1.National Research Center on Plant BiotechnologyIndian Agricultural Research InstituteNew DelhiIndia

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