Characterization of the U3 and U6 snRNA genes from wheat: U3 snRNA genes in monocot plants are transcribed by RNA polymerase III.
We have demonstrated recently that the genes encoding the U3 small nuclear RNA (snRNA) in dicot plants are transcribed by RNA polymerase III (pol III), and not RNA polymerase II (pol II) as in all other organisms studied to date. The U3 gene was the first example of a gene transcribed by different polymerases in different organisms. Based on phylogenetic arguments we proposed that a polymerase specificity change of the U3 snRNA gene promoter occurred during plant evolution. To map such an event we are examining the U3 gene polymerase specificity in other plant species. We report here the characterization of a U3 gene from wheat, a monocot plant. This gene contains the conserved promoter elements, USE and TATA, in a pol III-specific spacing seen also in a wheat U6 snRNA gene characterized in this report. Both the U3 and the U6 genes possess typical pol III termination signals but lack the cis element, responsible for 3'-end formation, found in all plant pol II-specific snRNA genes. In addition, expression of the U3 gene in transfected maize protoplasts is less sensitive to alpha-amanitin than a pol II-transcribed U2 gene. Based on these data we conclude that the wheat U3 gene is transcribed by pol III. This observation suggests that the postulated RNA polymerase specificity switch of the U3 gene took place prior to the divergence of angiosperm plants into monocots and dicots.
Marshallsay
C
C
Friedrich Miescher Institute, Basel, Switzerland.
Connelly
S
S
Filipowicz
W
W
D000546:Amanitins / Q000494:pharmacology; D001483:Base Sequence; D015870:Gene Expression / Q000187:drug effects; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009928:Organ Specificity / Q000235:genetics; D010802:Phylogeny; D011523:Protoplasts / Q000378:metabolism; D012320:RNA Polymerase III / Q000378:metabolism*; D012342:RNA, Small Nuclear / Q000235:genetics*; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D014162:Transfection; D014908:Triticum / Q000235:genetics*; D003313:Zea mays / Q000235:genetics
D000546:Amanitins; D012342:RNA, Small Nuclear; D012320:RNA Polymerase III