Characterization of transcription and processing from plasmids that use polIII and a yeast tRNA gene as promoter to transcribe promoter-deficient downstream DNA.
Transfer RNA (tRNA) with mutations affecting the internal promoters and thereby causing them to be nontranscribable by polIII (exemplified, e.g., by nematode tDNAPro with large insertions between the two internal promoters) could be transcribed by polIII both in vitro (yeast) and in vivo (oocytes) when cloned behind a yeast tRNAArg gene. PolIII initiated RNA synthesis could also proceed into a downstream structural gene normally read by polIII. the resultant yeast-nematode dimeric tRNA linked in front of mRNA sequences was recognized by processing enzymes to give mature tRNA. Thus, a yeast tRNA gene preceded by its 5' flank can function as a promoter for polIII transcription of any DNA, also of DNA coding for genes that otherwise could not have been expressed either in vitro or in vivo.
Otter
C A
CA
Department of Microbiology, University of Umeå, Sweden.
Edqvist
J
J
Stråby
K B
KB
D000818:Animals; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism*; D007763:Lac Operon / Q000235:genetics; D009348:Nematoda; D009693:Nucleic Acid Hybridization; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012343:RNA, Transfer / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014157:Transcription Factors / Q000378:metabolism*; D019474:Transcription Factors, TFIII; D014158:Transcription, Genetic
D014157:Transcription Factors; D019474:Transcription Factors, TFIII; C053256:transcription factor TFIIID; D004247:DNA; D012343:RNA, Transfer