Abstract
The Arabidopsis thaliana floral development protein AINTEGUMENTA (ANT) is a member of a large family of DNA binding proteins (AP2/ERF family) that control plant growth and development in response to developmental or environmental signals. Transcriptional activation and/or repression activities have been demonstrated for several members of this protein family. We have used fusions between ANT and the GAL4 DNA binding domain to identify an 80 amino acid sequence important for the transcriptional activation function of ANT. This region shows similarity to transcriptional activation domains in other proteins, as it is rich in Ser/Thr, Gln/Asn, and acidic amino acids. We also demonstrate that ANT can activate gene expression in Arabidopsis plants through binding to a DNA sequence corresponding to an in vitro determined ANT binding site. Finally, we show that ANT is nuclear localized and that the sequence KKKR (amino acids 252–255) is required for nuclear localization of the protein.
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Abbreviations
- Ler :
-
Landsberg erecta
- GBD:
-
GAL4 DNA binding domain
References
Baker SC, Robinson-Beers K, Villanueva JM, Gaiser JC, Gasser CS (1997) Interactions among genes regulating ovule development in Arabidopsis thaliana. Genetics 145:1109–1124
Bechtold N, Ellis J, Pelletier G (1993) In planta Agrobacterium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants. CR Acad Sci Ser III Sci Vie 316:1194–1199
Elliott RC, Betzner AS, Huttner E, Oakes MP, Tucker WQJ, Gerentes D, Perez P, Smyth DR (1996) AINTEGUMENTA, an APETALA2-like gene of Arabidopsis with pleiotropic roles in ovule development and floral organ growth. Plant Cell 8:155–168
Fujimoto SY, Ohta M, Usui A, Shinshi H, Ohme-Takagi M (2000) Arabidopsis ethylene-responsive element binding factors act as transcriptional activators or repressors of GCC box-mediated gene expression. Plant Cell 12:393–404
Gietz D, St. Jean A, Woods RA, Schiestl RH (1992) Improved method for high efficiency transformation of intact yeast cells. Nucleic Acids Res 20:1425
Johansen F-E, Prywes R (1993) Identification of transcriptional activation and inhibitory domains in serum response factor (SRF) by using GAL4-SRF constructs. Mol Cell Biol 13:4640–4647
Klucher KM, Chow H, Reiser L, Fischer RL (1996) The AINTEGUMENTA gene of Arabidopsis required for ovule and female gametophyte development is related to the floral homeotic gene APETALA2. Plant Cell 8:137–153
Krizek BA (1999) Ectopic expression of AINTEGUMENTA in Arabidopsis plants results in increased growth of floral organs. Dev Genet 25:224–236
Krizek BA (2003) AINTEGUMENTA utilizes a mode of DNA recognition distinct from that used by proteins containing a single AP2 domain. Nucleic Acids Res 31:1859–1868
Krizek BA, Meyerowitz EM (1996) The Arabidopsis genes APETALA3 and PISTILLATA are sufficient to specify the B class organ identity function. Development 122:11–22
Krizek BA, Prost V, Macias A (2000) AINTEGUMENTA promotes petal identity and acts as a negative regulator of AGAMOUS. Plant Cell 12:1357–1366
Liu L, White MJ, MacRae TH (1999) Transcription factors and their genes in higher plants: functional domains, evolution and regulation. Eur J Biochem 262:247–257
Liu Z, Franks RG, Klink VP (2000) Regulation of gynoecium marginal tissue formation by LEUNIG and AINTEGUMENTA. Plant Cell 12:1879–1891
Mizukami Y, Fischer RL (2000) Plant organ size control: AINTEGUMENTA regulates growth and cell numbers during organogenesis. Proc Natl Acad Sci USA 97:942–947
Nole-Wilson S, Krizek BA (2000) DNA binding properties of the Arabidopsis floral development protein AINTEGUMENTA. Nucleic Acids Res 28:4076–4082
Nole-Wilson S, Tranby T, Krizek BA (2005) AINTEGUMENTA-like (AIL) genes are expressed in young tissues and may specify meristematic or division-competent states. Plant Mol Biol 57:613–628
Ohta M, Matsui K, Hiratsu K, Shinshi H, Ohme-Takagi M (2001) Repression domains of class II ERF transcriptional repressors share an essential motif for active repression. Plant Cell 13:1959–1968
Rieu I, Bots M, Mariani C, Weterings KAP (2005) Isolation and expression analysis of a tobacco AINTEGUMENTA ortholog (NtANTL). Plant Cell Physiol 46:803–805
Schneitz K, Hulskamp M, Kopczak SD, Pruitt RE (1997) Dissection of sexual organ ontogenesis: a genetic analysis of ovule development in Arabidopsis thaliana. Development 124:1367–1376
Sridhar VV, Surendrarao A, Gonzalez D, Conlan RS, Liu Z (2004) Transcriptional repression of target genes by LEUNIG and SEUSS, two interacting regulatory proteins for Arabidopsis flower development. Proc Natl Acad Sci USA 101:11494–11499
Triezenberg SJ (1995) Structure and function of transcriptional activation domains. Curr Biol 5:190–196
Vergani P, Morandinin P, Soave C (1997) Complementation of a yeast Dpkc1 mutant by the Arabidopsis protein ANT. FEBS Lett 400:243–246
Wang Z, Triezenberg SJ, Thomashow MF, Stockinger EJ (2005) Multiple hydrophobic motifs in Arabidopsis CBF1 COOH-terminus provide functional redundancy in trans-activation. Plant Mol Biol 58:543–559
Acknowledgements
We thank Sandra Snyder for technical assistance with the gene gun experiments, Inhwan Hwang for the 326 GFP-3G plasmid, Natasha Grotz and Mary Lou Guerinot for advice on the leek bombardment experiments, and Deanna Smith for assistance with the GFP localization experiments. This work was supported by Department of Energy (Grant no. 98ER20312).
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Krizek, B.A., Sulli, C. Mapping sequences required for nuclear localization and the transcriptional activation function of the Arabidopsis protein AINTEGUMENTA. Planta 224, 612–621 (2006). https://doi.org/10.1007/s00425-006-0253-9
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DOI: https://doi.org/10.1007/s00425-006-0253-9