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Nucleic Acids Research, 2003, Vol. 31, No. 17 5003-5015
© 2003 Oxford University Press

Structural motifs in the RNA encoded by the early nodulation gene enod40 of soybean

Geneviève Girard1, Andreas Roussis1, Alexander P. Gultyaev1,2, Cornelis W. A. Pleij2 and Herman P. Spaink*,1

1 Institute of Biology, Leiden University, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands and 2 Institute of Chemistry, Leiden University, PO Box 9502, 2300 RA Leiden, The Netherlands

*To whom correspondence should be addressed. Tel: +31 71 527 5076; Fax: +31 71 527 5088; Email: spaink{at}rulbim.leidenuniv.nl
Present address:
Andreas Roussis, Center for Human and Clinical Genetics, Leiden University Medical Center, Wassenaarseweg 72, 2333 AL Leiden, The Netherlands

The plant gene enod40 is highly conserved among legumes and also present in various non-legume species. It is presumed to play a central regulatory role in the Rhizobium–legume interaction, being expressed well before the initiation of cortical cell divisions resulting in nodule formation. Two small peptides encoded by enod40 mRNA as well as its secondary structure have been shown to be key elements in the signalling processes underlying nodule organogenesis. Here results concerning the secondary structure of mRNA of enod40 in soybean are presented. This study combined a theoretical approach, involving structure prediction and comparison, as well as structure probing. Our study indicates five conserved domains in enod40 mRNA among numerous leguminous species. Structure comparison suggests that some domains are also conserved in non-leguminous species and that an additional domain exists that was found only in leguminous species developing indeterminate nodules. Enzymatic and chemical probing data support the structure for three of the domains, and partially for the remaining two. The rest of the molecule appears to be less structured. Some of the domains include motifs, such as U-containing internal loops and bulges, which seem to be conserved. Therefore, they might be involved in the regulatory role of enod40 RNA.


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