Translational and structural analysis of the shortest legume ENOD40 gene in Lupinus luteus.
Two early nodulin 40 (enod40) genes, ENOD40-1, the shortest legume ENOD40 gene, and ENOD40-2, were isolated from Lupinus luteus, a legume with indeterminate nodules. Both genes were expressed at similar levels during symbiosis with nitrogen-fixing bacteria. ENOD40 phylogeny clustered the L. luteus genes with legumes forming determinate nodules and revealed peptide similarities. The ENOD40-1 small ORF A fused to a reporter gene was efficiently expressed in plant cells, indicating that the start codon is recognized for translation. The ENOD40-1 RNA structure predicted based on Pb(II)-induced cleavage and modeling revealed four structurally conserved domains, an absence of domain 4 characteristic for legumes of indeterminate nodules, and interactions between the conserved region I and a region located upstream of domain 6. Domain 2 contains Mg(II) ion binding sites essential for organizing RNA secondary structure. The differences between L. luteus and Glycine max ENOD40 RNA models suggest the possibility of a switch between two structural states of ENOD40 transcript.
Podkowinski
Jan
J
Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznań, Poland. jpodkow@ibch.poznan.pl
Zmienko
Agnieszka
A
Florek
Blazena
B
Wojciechowski
Pawel
P
Rybarczyk
Agnieszka
A
Wrzesinski
Jan
J
Ciesiolka
Jerzy
J
Blazewicz
Jacek
J
Kondorosi
Adam
A
Crespi
Martin
M
Legocki
Andrzej
A
D015139:Blotting, Southern; D017930:Genes, Reporter; D031297:Lupinus / Q000235:genetics*; D009690:Nucleic Acid Conformation; D016366:Open Reading Frames; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics*; D014176:Protein Biosynthesis; D018749:RNA, Plant / Q000737:chemistry*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D010940:Plant Proteins; D018749:RNA, Plant