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The 8-nucleotide-long RNA : DNA hybrid is a primary stability determinant of the RNA polymerase II elongation complex

  1. Author:
    Kireeva, M. L.
    Komissarova, N.
    Waugh, D. S.
    Kashlev, M.
  2. Author Address

    Kashlev M NCI, Adv Biosci Labs Inc, Basic Res Program, Frederick Canc Res & Dev Ctr,NIH Bldg 539,Room 222 Frederick, MD 21702 USA NCI, Adv Biosci Labs Inc, Basic Res Program, Frederick Canc Res & Dev Ctr,NIH Frederick, MD 21702 USA
    1. Year: 2000
  1. Journal: Journal of Biological Chemistry
    1. 275
    2. 9
    3. Pages: 6530-6536
  2. Type of Article: Article
  1. Abstract:

    The sliding clamp model of transcription processivity, based on extensive studies of Escherichia coli RNA polymerase, suggests that formation of a stable elongation complex requires two distinct nucleic acid components: an 8-9-nt transcript-template hybrid, and a DNA duplex immediately downstream from the hybrid. Here, we address the minimal composition of the processive elongation complex in the eukaryotes by developing a method for promoter-independent assembly of functional elongation complex of S. cerevisiae RNA polymerase II from synthetic DNA and RNA oligonucleotides, me show that only one of the nucleic acid components, the 8-nt RNA: DNA hybrid, is necessary for the formation of a stable elongation complex with RNA polymerase II. The double-strand DNA upstream and downstream of the hybrid does not affect stability of the elongation complex. This finding reveals a significant difference in processivity determinants of RNA polymerase II and E. coli RNA polymerase. In addition, using the imperfect RNA:DNA hybrid disturbed by the mismatches in the RNA, we show that nontemplate DNA strand may reduce the elongation complex stability via the reduction of the RNA:DNA hybrid length. The structure of a "minimal stable" elongation complex suggests a key role of the RNA:DNA hybrid in RNA polymerase II processivity. [References: 47]

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