Mesh : Animals Base Sequence Cloning, Molecular DNA Restriction Enzymes DNA-Directed RNA Polymerases / metabolism Genes Promoter Regions, Genetic RNA Polymerase II / metabolism RNA Polymerase III / metabolism RNA, Small Nuclear / genetics Repetitive Sequences, Nucleic Acid Transcription, Genetic Xenopus

来  源:   DOI:10.1093/nar/15.6.2463   PDF(Sci-hub)

Abstract:
We have cloned and sequenced a 977bp DNA fragment, pXTU6-2, that represents the transcription unit for a Xenopus tropicalis U6 RNA gene. This basic repeating unit is reiterated ca.500-fold per haploid genome. Oocyte injections of pXTU6-2 led to the transcription of a mature-sized U6 RNA that, however, lacked internal 2\'-O-methylations. These posttranscriptional modifications of U6 RNA might be cytoplasmic and could require its association with U4 RNA to be accomplished. The low alpha- amanitin sensitivity of U6 RNA synthesis in oocytes suggested that U6 RNA is transcribed by RNA polymerase III, consistent with features of the U6 RNA molecule which also contains a Box A- like intragenic control region. Inspection of X. tropicalis, mouse and human U6 DNA upstream sequences revealed the presence of a TATA box as well as of the proximal and enhancer (octamer motif) elements contained in snRNA genes transcribed by RNA polymerase II. We propose that U6 RNAs are synthesized by a specialized transcription complex consisting of RNA polymerase III and transcription factors, some of which are very likely shared with RNA polymerase II promoters.
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