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J. Biol. Chem., Vol. 278, Issue 38, 36396-36402, September 19, 2003

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Functional Analysis of RF2a, a Rice Transcription Factor^*

Shunhong Dai  , Silvana Petruccelli  ¶ , Maria Isabel Ordiz  ||, Zhihong Zhang , Shouyi Chen  and Roger N. Beachy  **

From the Donald Danforth Plant Science Center, St. Louis, Missouri 63132 and the Institute of Genetics, Chinese Academy of Sciences, Beijing 100101, China

RF2a is a bZIP transcription factor that regulates expression of the promoter of rice tungro bacilliform badnavirus. RF2a is predicted to include three domains that contribute to its function. The results of transient assays with mutants of RF2a from which one or more domains were removed demonstrated that the acidic domain was essential for the activation of gene expression, although the proline-rich and glutamine-rich domains each played a role in this function. Studies using fusion proteins of different functional domains of RF2a with the 2C7 synthetic zinc finger DNA-binding domain showed that the acidic region is a relatively strong activation domain, the function of which is dependent on the context in which the domain is placed. Data from transgenic plants further supported the conclusion that the acidic domain was important for maintaining the biological function of RF2a. RF2a and TBP (TATA-binding protein) synergistically activate transcription in vitro (Zhu, Q., Ordiz, M. I., Dabi, T., Beachy, R. N., and Lamb, C. (2002) Plant Cell 14, 795–803). In vitro and in vivo assays showed that RF2a interacts with TBP through the glutamine-rich domain but not the acidic domain. Functional analysis of such interactions indicates that the acidic domain activates transcription through mechanisms other than via the direct recruitment of TBP.

Received for publication, May 8, 2003 , and in revised form, July 8, 2003.

^* This work was supported by Department of Energy Grant DE-FG02-99ER20355 (to R. N. B.) and by funds from the Donald Danforth Plant Science Center. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ Present address: Centro de Investigación y Desarrollo en Criotecnología de Alimentos, 47 y 116, 1900 La Plata, Argentina.

Supported in part by Argentina National Council Research (CONICET).

^|| Supported in part by Fundación para el Fomento de la Investigación Científica Aplicada y la Tecnología (Spain).

^** To whom correspondence should be addressed: Donald Danforth Plant Science Center, 975 North Warson Rd., St. Louis, MO 63132. E-mail: rnbeachy{at}danforthcenter.org.

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