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. 1998 Jun;149(2):479–490. doi: 10.1093/genetics/149.2.479

Evidence for a role for AtMYB2 in the induction of the Arabidopsis alcohol dehydrogenase gene (ADH1) by low oxygen.

F U Hoeren 1, R Dolferus 1, Y Wu 1, W J Peacock 1, E S Dennis 1
PMCID: PMC1460183  PMID: 9611167

Abstract

The transcription factor AtMYB2 binds to two sequence motifs in the promoter of the Arabidopsis ADH1 gene. The binding to the GT-motif (5'-TGGTTT-3') is essential for induction of ADH1 by low oxygen, while binding to the second motif, MBS-2, is not essential for induction. We show that AtMYB2 is induced by hypoxia with kinetics compatible with a role in the regulation of ADH1. Like ADH1, AtMYB2 has root-limited expression. When driven by a constitutive promoter, AtMYB2 is able to transactivate ADH1 expression in transient assays in both Arabidopsis and Nicotiana plumbaginifolia protoplasts, and in particle bombardment of Pisum sativum leaves. Mutation of the GT-motif abolished binding of AtMYB2 and caused loss of activity of the ADH1 promoter in both transient assays and transgenic Arabidopsis plants. These results are consistent with AtMYB2 being a key regulatory factor in the induction of the ADH1 promoter by low oxygen.

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Selected References

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  1. Abe H., Yamaguchi-Shinozaki K., Urao T., Iwasaki T., Hosokawa D., Shinozaki K. Role of arabidopsis MYC and MYB homologs in drought- and abscisic acid-regulated gene expression. Plant Cell. 1997 Oct;9(10):1859–1868. doi: 10.1105/tpc.9.10.1859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Abel S., Theologis A. Transient transformation of Arabidopsis leaf protoplasts: a versatile experimental system to study gene expression. Plant J. 1994 Mar;5(3):421–427. doi: 10.1111/j.1365-313x.1994.00421.x. [DOI] [PubMed] [Google Scholar]
  3. Bailey-Serres J., Freeling M. Hypoxic stress-induced changes in ribosomes of maize seedling roots. Plant Physiol. 1990 Nov;94(3):1237–1243. doi: 10.1104/pp.94.3.1237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Berberich T., Kusano T. Cycloheximide induces a subset of low temperature-inducible genes in maize. Mol Gen Genet. 1997 Apr 16;254(3):275–283. doi: 10.1007/s004380050416. [DOI] [PubMed] [Google Scholar]
  5. Bevan M. Binary Agrobacterium vectors for plant transformation. Nucleic Acids Res. 1984 Nov 26;12(22):8711–8721. doi: 10.1093/nar/12.22.8711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Burk O., Mink S., Ringwald M., Klempnauer K. H. Synergistic activation of the chicken mim-1 gene by v-myb and C/EBP transcription factors. EMBO J. 1993 May;12(5):2027–2038. doi: 10.1002/j.1460-2075.1993.tb05852.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burke T. J., Callis J., Vierstra R. D. Characterization of a polyubiquitin gene from Arabidopsis thaliana. Mol Gen Genet. 1988 Aug;213(2-3):435–443. doi: 10.1007/BF00339613. [DOI] [PubMed] [Google Scholar]
  8. Damm B., Schmidt R., Willmitzer L. Efficient transformation of Arabidopsis thaliana using direct gene transfer to protoplasts. Mol Gen Genet. 1989 May;217(1):6–12. doi: 10.1007/BF00330935. [DOI] [PubMed] [Google Scholar]
  9. Dennis E. S., Gerlach W. L., Walker J. C., Lavin M., Peacock W. J. Anaerobically regulated aldolase gene of maize. A chimaeric origin? J Mol Biol. 1988 Aug 20;202(4):759–767. doi: 10.1016/0022-2836(88)90556-6. [DOI] [PubMed] [Google Scholar]
  10. Dennis E. S., Sachs M. M., Gerlach W. L., Finnegan E. J., Peacock W. J. Molecular analysis of the alcohol dehydrogenase 2 (Adh2) gene of maize. Nucleic Acids Res. 1985 Feb 11;13(3):727–743. doi: 10.1093/nar/13.3.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dolferus R., Jacobs M., Peacock W. J., Dennis E. S. Differential interactions of promoter elements in stress responses of the Arabidopsis Adh gene. Plant Physiol. 1994 Aug;105(4):1075–1087. doi: 10.1104/pp.105.4.1075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ferl R. J., Nick H. S. In vivo detection of regulatory factor binding sites in the 5' flanking region of maize Adh1. J Biol Chem. 1987 Jun 15;262(17):7947–7950. [PubMed] [Google Scholar]
  13. Gleave A. P. A versatile binary vector system with a T-DNA organisational structure conducive to efficient integration of cloned DNA into the plant genome. Plant Mol Biol. 1992 Dec;20(6):1203–1207. doi: 10.1007/BF00028910. [DOI] [PubMed] [Google Scholar]
  14. Goff S. A., Cone K. C., Chandler V. L. Functional analysis of the transcriptional activator encoded by the maize B gene: evidence for a direct functional interaction between two classes of regulatory proteins. Genes Dev. 1992 May;6(5):864–875. doi: 10.1101/gad.6.5.864. [DOI] [PubMed] [Google Scholar]
  15. Grotewold E., Athma P., Peterson T. Alternatively spliced products of the maize P gene encode proteins with homology to the DNA-binding domain of myb-like transcription factors. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4587–4591. doi: 10.1073/pnas.88.11.4587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Grotewold E., Drummond B. J., Bowen B., Peterson T. The myb-homologous P gene controls phlobaphene pigmentation in maize floral organs by directly activating a flavonoid biosynthetic gene subset. Cell. 1994 Feb 11;76(3):543–553. doi: 10.1016/0092-8674(94)90117-1. [DOI] [PubMed] [Google Scholar]
  17. Gubler F., Kalla R., Roberts J. K., Jacobsen J. V. Gibberellin-regulated expression of a myb gene in barley aleurone cells: evidence for Myb transactivation of a high-pI alpha-amylase gene promoter. Plant Cell. 1995 Nov;7(11):1879–1891. doi: 10.1105/tpc.7.11.1879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hossain M. A., Huq E., Grover A., Dennis E. S., Peacock W. J., Hodges T. K. Characterization of pyruvate decarboxylase genes from rice. Plant Mol Biol. 1996 Jul;31(4):761–770. doi: 10.1007/BF00019464. [DOI] [PubMed] [Google Scholar]
  19. Köhler U., Liaud M. F., Mendel R. R., Cerff R., Hehl R. The maize GapC4 promoter confers anaerobic reporter gene expression and shows homology to the maize anthocyanin regulatory locus C1. Plant Mol Biol. 1995 Dec;29(6):1293–1298. doi: 10.1007/BF00020469. [DOI] [PubMed] [Google Scholar]
  20. Lazo G. R., Stein P. A., Ludwig R. A. A DNA transformation-competent Arabidopsis genomic library in Agrobacterium. Biotechnology (N Y) 1991 Oct;9(10):963–967. doi: 10.1038/nbt1091-963. [DOI] [PubMed] [Google Scholar]
  21. Llewellyn D. J., Finnegan E. J., Ellis J. G., Dennis E. S., Peacock W. J. Structure and expression of an alcohol dehydrogenase 1 gene from Pisum sativum (cv. "Greenfeast"). J Mol Biol. 1987 May 5;195(1):115–123. doi: 10.1016/0022-2836(87)90331-7. [DOI] [PubMed] [Google Scholar]
  22. Lång V., Palva E. T. The expression of a rab-related gene, rab18, is induced by abscisic acid during the cold acclimation process of Arabidopsis thaliana (L.) Heynh. Plant Mol Biol. 1992 Dec;20(5):951–962. doi: 10.1007/BF00027165. [DOI] [PubMed] [Google Scholar]
  23. Millar A. A., Dennis E. S. The alcohol dehydrogenase genes of cotton. Plant Mol Biol. 1996 Jul;31(4):897–904. doi: 10.1007/BF00019476. [DOI] [PubMed] [Google Scholar]
  24. Olive M. R., Walker J. C., Singh K., Dennis E. S., Peacock W. J. Functional properties of the anaerobic responsive element of the maize Adh1 gene. Plant Mol Biol. 1990 Oct;15(4):593–604. doi: 10.1007/BF00017834. [DOI] [PubMed] [Google Scholar]
  25. Roth B. A., Goff S. A., Klein T. M., Fromm M. E. C1- and R-dependent expression of the maize Bz1 gene requires sequences with homology to mammalian myb and myc binding sites. Plant Cell. 1991 Mar;3(3):317–325. doi: 10.1105/tpc.3.3.317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sablowski R. W., Moyano E., Culianez-Macia F. A., Schuch W., Martin C., Bevan M. A flower-specific Myb protein activates transcription of phenylpropanoid biosynthetic genes. EMBO J. 1994 Jan 1;13(1):128–137. doi: 10.1002/j.1460-2075.1994.tb06242.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sachs M. M., Freeling M., Okimoto R. The anaerobic proteins of maize. Cell. 1980 Jul;20(3):761–767. doi: 10.1016/0092-8674(80)90322-0. [DOI] [PubMed] [Google Scholar]
  28. Shen Q., Uknes S. J., Ho T. H. Hormone response complex in a novel abscisic acid and cycloheximide-inducible barley gene. J Biol Chem. 1993 Nov 5;268(31):23652–23660. [PubMed] [Google Scholar]
  29. Shen Q., Zhang P., Ho T. H. Modular nature of abscisic acid (ABA) response complexes: composite promoter units that are necessary and sufficient for ABA induction of gene expression in barley. Plant Cell. 1996 Jul;8(7):1107–1119. doi: 10.1105/tpc.8.7.1107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Solano R., Nieto C., Avila J., Cañas L., Diaz I., Paz-Ares J. Dual DNA binding specificity of a petal epidermis-specific MYB transcription factor (MYB.Ph3) from Petunia hybrida. EMBO J. 1995 Apr 18;14(8):1773–1784. doi: 10.1002/j.1460-2075.1995.tb07166.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tice-Baldwin K., Fink G. R., Arndt K. T. BAS1 has a Myb motif and activates HIS4 transcription only in combination with BAS2. Science. 1989 Nov 17;246(4932):931–935. doi: 10.1126/science.2683089. [DOI] [PubMed] [Google Scholar]
  32. Urao T., Noji M., Yamaguchi-Shinozaki K., Shinozaki K. A transcriptional activation domain of ATMYB2, a drought-inducible Arabidopsis Myb-related protein. Plant J. 1996 Dec;10(6):1145–1148. doi: 10.1046/j.1365-313x.1996.10061145.x. [DOI] [PubMed] [Google Scholar]
  33. Urao T., Yamaguchi-Shinozaki K., Urao S., Shinozaki K. An Arabidopsis myb homolog is induced by dehydration stress and its gene product binds to the conserved MYB recognition sequence. Plant Cell. 1993 Nov;5(11):1529–1539. doi: 10.1105/tpc.5.11.1529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Valvekens D., Van Montagu M., Van Lijsebettens M. Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5536–5540. doi: 10.1073/pnas.85.15.5536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wang Z. Y., Kenigsbuch D., Sun L., Harel E., Ong M. S., Tobin E. M. A Myb-related transcription factor is involved in the phytochrome regulation of an Arabidopsis Lhcb gene. Plant Cell. 1997 Apr;9(4):491–507. doi: 10.1105/tpc.9.4.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Werr W., Frommer W. B., Maas C., Starlinger P. Structure of the sucrose synthase gene on chromosome 9 of Zea mays L. EMBO J. 1985 Jun;4(6):1373–1380. doi: 10.1002/j.1460-2075.1985.tb03789.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Weston K. Extension of the DNA binding consensus of the chicken c-Myb and v-Myb proteins. Nucleic Acids Res. 1992 Jun 25;20(12):3043–3049. doi: 10.1093/nar/20.12.3043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wolyn D. J., Jelenkovic G. Nucleotide sequence of an alcohol dehydrogenase gene in octoploid strawberry (Fragaria x Ananassa Duch). Plant Mol Biol. 1990 May;14(5):855–857. doi: 10.1007/BF00016518. [DOI] [PubMed] [Google Scholar]
  39. Xie Y., Wu R. Molecular analysis of an alcohol dehydrogenase-encoding genomic clone (adh2) from rice. Gene. 1990 Mar 15;87(2):185–191. doi: 10.1016/0378-1119(90)90300-g. [DOI] [PubMed] [Google Scholar]
  40. de Bruxelles G. L., Peacock W. J., Dennis E. S., Dolferus R. Abscisic acid induces the alcohol dehydrogenase gene in Arabidopsis. Plant Physiol. 1996 Jun;111(2):381–391. doi: 10.1104/pp.111.2.381. [DOI] [PMC free article] [PubMed] [Google Scholar]

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