myb7 mRNA is present in rice in spliced and unspliced forms, splicing being enhanced by anoxia. The protein (Mybleu) encoded by the unspliced mRNA is composed of an incomplete Myb domain followed by a leucine zipper; however, it lacks canonical sequences for DNA binding, transcriptional activation, and nuclear localization. We show here that in transiently transformed tobacco protoplasts, Mybleu is able to enhance the transcriptional activity of the maize leucine zipper Opaque2 on its target b32 promoter. The Mybleu transactivation effect is strictly dependent on the presence of Opaque2 and is driven by Mybleu-Opaque2 heterodimers. Mybleu is located in the nucleus, both in rice and in transformed tobacco protoplasts. In rice, the protein is expressed in regions corresponding to undifferentiated cells of roots and coleoptiles. Therefore, myb7 mRNA encodes, depending on its splicing, two transcription factors belonging to separate classes. One of them, Mybleu, has novel structural characteristics, suggesting the existence of new mechanisms acting in the activation of transcription.Mybs are a family of transcription factors widely represented in viruses, insects, mammals, and plants. The common feature of Myb factors is the presence of a conserved domain consisting of imperfect repeats of 50 -53 amino acids, called "tryptophan clusters" because of the highly conserved tryptophan residues involved in stabilizing the structure of the DNA binding domain (1). In plants, myb genes are present as large families (6 -100 members) involved in the control of a wide range of biochemical pathways (2-6), including responses to biotic and abiotic stresses (7-13).Screening a cDNA library from anaerobically grown rice coleoptiles, we isolated myb7, a cDNA derived from a Mybencoding unspliced mRNA (8). Some features of myb7 sequence indicate that it may be post-transcriptionally regulated; in particular, two unspliced introns are present in positions that are conserved with respect to other plant myb genes. Both spliced and unspliced myb7 mRNAs are present in vivo (7).In aerobically grown rice roots, we observed a higher proportion of unspliced myb7 RNA, with respect to the spliced form. The spliced form is instead predominant in rice roots during anoxia, a stress situation that generally inhibits splicing (14). These data suggest that post-transcriptional regulation controls the ratio between myb7 unspliced and spliced forms (7). This hypothesis is supported by the observation that the 5Ј region of the first intron can direct the synthesis of an in-frame leucine zipper, a functional domain present in several transcription factors (15-18).The putative polypeptide encoded by the unspliced myb7 mRNA consists of an incomplete Myb domain followed by the leucine zipper: we therefore named it Mybleu. Mybleu does not have the characteristics of a functional transcription factor. Indeed, it should be unable to bind DNA because of the presence of an incomplete Myb repeat. Moreover, neither a putative transcriptional activator region nor a c...
We have isolated two overlapping cDNAs coding for a MYB-related protein expressed in aerobic and anaerobic rice (Oryza sativa) roots and coleoptiles. Analysis of their sequences reveals some peculiar features, suggesting the presence of post-transcriptional regulation events: an upstream ORF, two unspliced introns and a putative leucine zipper in the ORF coded by the unspliced RNA. Transient expression in protoplasts indicates that the upstream ORF inhibits expression of a downstream coding sequence. Finally, we demonstrated that anoxia, in roots, increases the ratio between the spliced and the unspliced mRNA and affects the expression of other myb-related genes.
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