Structural analysis of ternary complexes of vaccinia RNA polymerase Journal Article
| Authors: | Hagler, J.; Shuman, S. |
| Article Title: | Structural analysis of ternary complexes of vaccinia RNA polymerase |
| Abstract: | The structure of the elongation complex of vaccinia RNA polymerase halted at discrete template positions was examined by DNase I footprinting. The leading edge of the footprint bore a constant relationship to the catalytic template position, being 22-24 nucleotides (nt) in advance on the nontemplate strand and 17 nt on the template strand. DNase hypersensitivity of the nontemplate strand at the leading edge suggested that the DNA might be distorted as it entered the polymerase molecule. The region of DNA unwinding at the transcription bubble extended at least 12 nt 5′ from the catalytic center, as indicated by the reactivity of adenosine residues to diethylpyrocarbonate. Cu-phenanthroline-hypersensitive sites located 13 nt 5′ and 4 nt 3′ of the growing point appeared to demarcate the margins of the bubble. Strand asymmetry of chemical modification within the bubble was consistent with an RNA-DNA hybrid of no more than 10 base pairs. |
| Keywords: | transcription initiation; transcription factor; transcription, genetic; transcription factors; molecular sequence data; messenger rna; adenosine; nucleotide sequence; vaccinia virus; base sequence; base pairing; enzyme structure; enzyme assay; protein dna interaction; enzyme active site; dna-directed rna polymerases; dna denaturation; rna polymerase; oligodeoxyribonucleotides; elongation factor; vaccinia; dna rna hybridization; dna template; dna protein complex; templates, genetic; dna footprinting; restriction mapping; promoter regions (genetics); deoxyribonuclease i; priority journal; article; macromolecular systems; footprint; mrna elongation; transcription bubble; 1,10 phenanthroline copper; diethyl pyrocarbonate |
| Journal Title: | Proceedings of the National Academy of Sciences of the United States of America |
| Volume: | 89 |
| Issue: | 21 |
| ISSN: | 0027-8424 |
| Publisher: | National Academy of Sciences |
| Date Published: | 1992-11-01 |
| Start Page: | 10099 |
| End Page: | 10103 |
| Language: | English |
| DOI: | 10.1073/pnas.89.21.10099 |
| PUBMED: | 1438198 |
| PROVIDER: | scopus |
| PMCID: | PMC50285 |
| DOI/URL: | |
| Notes: | Source: Scopus |
