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Molecular and Cellular Biology, January 2001, p. 354-366, Vol. 21, No. 1
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.1.354-366.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Translational and Structural Requirements of the Early Nodulin Gene enod40, a Short-Open Reading Frame-Containing RNA, for Elicitation of a Cell-Specific Growth Response in the Alfalfa Root Cortex

Carolina Sousa,1,dagger Christina Johansson,1,Dagger Celine Charon,1 Hamid Manyani,1 Christof Sautter,2 Adam Kondorosi,1,3,* and Martin Crespi1

Institut des Sciences Végétales, Centre National de la Recherche Scientifique, F-91198 Gif-sur-Yvette Cedex, France1; Institute of Plant Sciences, Swiss Federal Institute of Technology, CH-8092 Zürich, Switzerland2; and Institute of Genetics, Biological Research Center, Hungarian Academy of Sciences, H-6701 Szeged, Hungary3

Received 29 June 2000/Returned for modification 3 August 2000/Accepted 3 October 2000

A diversity of mRNAs containing only short open reading frames (sORF-RNAs; encoding less than 30 amino acids) have been shown to be induced in growth and differentiation processes. The early nodulin gene enod40, coding for a 0.7-kb sORF-RNA, is expressed in the nodule primordium developing in the root cortex of leguminous plants after infection by symbiotic bacteria. Ballistic microtargeting of this gene into Medicago roots induced division of cortical cells. Translation of two sORFs (I and II, 13 and 27 amino acids, respectively) present in the conserved 5' and 3' regions of enod40 was required for this biological activity. These sORFs may be translated in roots via a reinitiation mechanism. In vitro translation products starting from the ATG of sORF I were detectable by mutating enod40 to yield peptides larger than 38 amino acids. Deletion of a Medicago truncatula enod40 region between the sORFs, spanning a predicted RNA structure, did not affect their translation but resulted in significantly decreased biological activity. Our data reveal a complex regulation of enod40 action, pointing to a role of sORF-encoded peptides and structured RNA signals in developmental processes involving sORF-RNAs.

* Corresponding author. Mailing address: Institut des Sciences Végétales, Centre National de la Recherche Scientifique, F-91198 Gif-sur-Yvette, France. Phone: 33-1-69823696. Fax: 33-1-69823695. E-mail: Adam.Kondorosi{at}isv.cnrs-gif.fr.

dagger Present address: Department of Microbiology and Parasitology, University of Seville, 41080 Seville, Spain.

Dagger Present address: Department of Molecular and Structural Biology, Aarhus University, DK-8000 Aarhus, Denmark.

Molecular and Cellular Biology, January 2001, p. 354-366, Vol. 21, No. 1
0270-7306/01/$04.00+0 DOI: 10.1128/MCB.21.1.354-366.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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