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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,
Christina
Johansson,1,
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.

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

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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