Yeast class III gene transcription factors and homologous RNA polymerase III form ternary transcription complexes stable to disruption by N-lauroyl-sarcosine (sarcosyl).
Yeast Class III gene transcription factors and RNA polymerase III were used to form ternary transcription complexes on a tRNASer gene in vitro under UTP-limiting transcription conditions. These ternary transcription complexes were composed of template DNA, proteins, and RNA. We have shown that the RNAs contained in these complexes represented specifically initiated nascent pre-tRNASer transcripts. These nascent RNAs could be very efficiently elongated to full-length pre-tRNASer molecules, even in the presence of the ionic detergent sarcosyl. Partial purification (greater than 100-fold) of these sarcosyl-resistant ternary transcription complexes could be achieved in a single step via sucrose gradient sedimentation. Comparable sarcosyl-resistant ternary transcription complexes could not be formed using purified yeast RNA polymerase III as the only protein component of the complex.
M S
MS
Klekamp
P A
PA
Weil
D001483:Base Sequence; D002499:Centrifugation, Density Gradient; D002852:Chromatography, Ion Exchange; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D003902:Detergents / Q000494:pharmacology*; D008970:Molecular Weight; D009694:Nucleic Acid Precursors / Q000032:analysis; D011485:Protein Binding / Q000187:drug effects; D012320:RNA Polymerase III / Q000378:metabolism*; D012322:RNA Precursors; D012343:RNA, Transfer / Q000032:analysis; D012346:RNA, Transfer, Amino Acyl / Q000032:analysis; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D012521:Sarcosine / Q000031:analogs & derivatives* / Q000494:pharmacology; D013501:Surface-Active Agents / Q000494:pharmacology*; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects*
D003902:Detergents; D009694:Nucleic Acid Precursors; D012322:RNA Precursors; D012346:RNA, Transfer, Amino Acyl; D013501:Surface-Active Agents; D014157:Transcription Factors; C025231:sarkosyl; D012343:RNA, Transfer; D012321:DNA-Directed RNA Polymerases; D012320:RNA Polymerase III; D012521:Sarcosine