Yeast class III gene transcription factors and homologous RNA polymerase III form ternary transcription complexes stable to disruption by N-lauroyl-sarcosine (sarcosyl).

Klekamp MS, Weil PA
Arch Biochem Biophys. 1986 246 (2): 783-800

PMID: 2423033 · DOI:10.1016/0003-9861(86)90335-8

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.

MeSH Terms (17)

Base Sequence Centrifugation, Density Gradient Chromatography, Ion Exchange Detergents DNA-Directed RNA Polymerases Molecular Weight Nucleic Acid Precursors Protein Binding RNA, Transfer RNA, Transfer, Amino Acyl RNA Polymerase III RNA Precursors Saccharomyces cerevisiae Sarcosine Surface-Active Agents Transcription, Genetic Transcription Factors

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