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J. Biol. Chem., Vol. 279, Issue 2, 1468-1473, January 9, 2004

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An Arabidopsis RNA Lariat Debranching Enzyme Is Essential for Embryogenesis^*

Huai Wang, Kristine Hill, and Sharyn E. Perry

From the Department of Agronomy, University of Kentucky, Lexington, Kentucky 40546-0312

An embryo-defective mutant of Arabidopsis thaliana was isolated that arrests development at a variety of stages, from as early as the globular stage of embryogenesis to as late as formation of an abnormal bent cotyledon stage embryo. Defects in the suspensor, a normally transient structure derived from the fertilized egg, were often associated with the arrested embryo. The lesion was within a gene encoding a protein with domains characteristic of lariat debranching enzymes, which has been named AtDBR1 (for Arabidopsis thaliana Debranching enzyme 1). Cleavage of the 2'-5'-phosphodiester bond found in excised intron lariats ("debranching") is essential for turnover of intronic sequences as well as generation of some small nucleolar RNAs. The mutation within AtDBR1 was confirmed by complementation as being responsible for the embryo-lethal phenotype, and the activity of the encoded protein in cleavage of 2'-5'-phosphodiester bonds was verified using an in vitro debranching assay.

Received for publication, August 18, 2003 , and in revised form, October 14, 2003.

^* This work was supported by National Science Foundation Award IBN-9984274 (to S. E. P.), the United States Department of Agriculture, NRICGP, Plant Genetic Mechanisms Award 2002-35301-12026 (to S. E. P.), and by the University of Kentucky. This is paper number 03-06-094) from the Kentucky Agricultural Experiment Station. 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.

To whom correspondence should be addressed: Dept. of Agronomy, University of Kentucky, 1405 Veterans Dr., 307 Plant Science Bldg., Lexington, KY 40546-0312. Tel.: 859-257-5020 (ext. 80732); Fax: 859-257-7125; E-mail: sperr2{at}uky.edu.

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