Atypical Splicing Accompanied by Skipping Conserved Micro-Exons Produces Unique WRINKLED1, An AP2 Domain Transcription Factor in Rice Plants

WRINKLED1 (WRI1), an AP2 domain transcription factor, is a master regulator of oil synthesis in plant seeds. Its closely related proteins (WRIs) are also involved in regulating the synthesis of fatty acids, which play a role in producing oils, membranes, and other important components in plants. We found two WRI1 genes, OsWRI1-1 and OsWRI1-2, and two additional WRI1 homologs, OsWRI3 and OsWRI4, in the rice genome. OsWRI1 was ubiquitously expressed in rice plants, including developing seeds. However, OsWRI3 was only significantly expressed in the leaf blade and OsWRI4 was not expressed at all. OsWRI1-1 contains amino acid sequence GCL instead of VYL, which is encoded by an independent 9-bp micro-exon that is conserved in many plant species. We found that the GCL sequence was produced by an atypical splicing accompanied by skipping of the micro-exon. Furthermore, OsWRI1-1 highly activates the transcription of the promoter for the biotin carboxyl transferase 2 gene in Arabidopsis, but its activity was reduced by amino acid replacement or deletion of the GCL sequence in a transient assay using Arabidopsis cells. Our results indicated that atypical splicing produced unique WRI1 in rice plants.