title: RNA polymerase III accurately initiates transcription from RNA polymerase II promoters in vitro.
abstract: In eukaryotes, there are three major RNA polymerases (Pol) in the nucleus, which are commonly described as transcribing non-overlapping subsets of genes. Structural studies have highlighted a conserved core shared among all three transcription systems. Initiation of human Pol III from TATA box-containing Pol II promoters under conditions with impaired Pol II transcription activity have been described previously. RNA polymerase III and Pol II were found to co-localize at the promoters of the c-myc gene and the RPPH1 sRNA in vivo. Here, I report that Pol III can, like Pol II, initiate transcription from most tested Pol II core promoters when assayed with crude human nuclear extracts (HSK, SNF, or Dignam). Both polymerases often initiate from the same transcription start site, and depend on a TATA box or AT-rich region but not the downstream promoter element (DPE) or the motif ten element (MTE). Moderate (∼2-fold) changes in the ratio of DNA template to nuclear extract were sufficient to change Pol II-mediated transcription to a mixture of Pol II- and Pol III-, or to a solely Pol III-dependent initiation of transcription from Pol II promoters. Polymerase specificity is thus not fixed but a variable that depends on the properties of the promoter and the transcription conditions. These findings provide functional evidence for a close similarity between the Pol II and Pol III transcription complexes, and additionally explain previous controversies in the literature.
authors.affiliation: From the Section of Molecular Biology, Division of Biological Sciences, University of California at San Diego, La Jolla, California 92093 sduttke@ucsd.edu.
authors.forename: Sascha H C
authors.initials: SH
authors.lastname: Duttke
mesh_terms: D020856:AT Rich Sequence; D004247:DNA / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D011401:Promoter Regions, Genetic; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D012319:RNA Polymerase II / Q000235:genetics* / Q000378:metabolism; D012320:RNA Polymerase III / Q000378:metabolism*; D013379:Substrate Specificity; D016385:TATA Box; D061785:Transcription Initiation, Genetic
chemical_list: D012313:RNA; D004247:DNA; D012319:RNA Polymerase II; D012320:RNA Polymerase III
keywords: Gene Regulation; Polymerase Specificity; RNA Polymerase II; RNA Polymerase III; RNA Synthesis; TATA Box; TBP; Transcription; α-Amanitin