Elongation complexes of thermus thermophilus RNA polymerase that possess distinct translocation conformations

Ekaterina Kashkina, Michael Anikin, Tahir H Tahirov, Sergei N. Kochetkov, Dmitry G. Vassylyev, Dmitry Temiakov

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

We have characterized elongation complexes (ECs) of RNA polymerase from the extremely thermophilic bacterium, Thermus thermophilus. We found that complexes assembled on nucleic acid scaffolds are transcriptionally competent at high temperature (50 - 80°C) and, depending upon the organization of the scaffold, possess distinct translocation conformations. ECs assembled on scaffolds with a 9 bp RNA:DNA hybrid are highly stable, resistant to pyrophosphorolysis, and are in the posttranslocated state. ECs with an RNA:DNA hybrid longer or shorter than 9 bp appear to be in a pretranslocated state, as evidenced by their sensitivity to pyrophosphorolysis, GreA-induced cleavage, and exonuclease footprinting. Both pretranslocated (8 bp RNA:DNA hybrid) and posttranslocated (9 bp RNA:DNA hybrid) complexes were crystallized in distinct crystal forms, supporting the homogeneity of the conformational states in these complexes. Crystals of a posttranslocated complex were used to collect diffraction data at atomic resolution.

Original languageEnglish (US)
Pages (from-to)4036-4045
Number of pages10
JournalNucleic acids research
Volume34
Issue number14
DOIs
StatePublished - Sep 21 2006

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Thermus thermophilus
DNA-Directed RNA Polymerases
RNA
DNA
Exonucleases
Nucleic Acids
Bacteria
Temperature

ASJC Scopus subject areas

  • Genetics

Cite this

Elongation complexes of thermus thermophilus RNA polymerase that possess distinct translocation conformations. / Kashkina, Ekaterina; Anikin, Michael; Tahirov, Tahir H; Kochetkov, Sergei N.; Vassylyev, Dmitry G.; Temiakov, Dmitry.

In: Nucleic acids research, Vol. 34, No. 14, 21.09.2006, p. 4036-4045.

Research output: Contribution to journalArticle

Kashkina, Ekaterina ; Anikin, Michael ; Tahirov, Tahir H ; Kochetkov, Sergei N. ; Vassylyev, Dmitry G. ; Temiakov, Dmitry. / Elongation complexes of thermus thermophilus RNA polymerase that possess distinct translocation conformations. In: Nucleic acids research. 2006 ; Vol. 34, No. 14. pp. 4036-4045.
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