DOI: 10.1073/pnas.95.25.14699. Functional topography of nascent RNA in elongation intermediates of RNA polymerase

Functional topography of nascent RNA in elongation intermediates of RNA polymerase

10.1073/pnas.95.25.14699

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

To determine the dynamics of transcript extrusion from Escherichia coli RNA polymerase (RNAP), we used degradation of the RNA by RNases T1 and A in a series of consecutive elongation complexes (ECs). In intact ECs, even extremely high doses of the RNases were unable to cut the RNA closer than 14–16 nt from the 3′ end. Our results prove that all of the cuts detected within the 14-nt zone are derived from the EC that is denatured during inactivation of the RNases. The protected zone monotonously translocates along the RNA after addition of new nucleotides to the transcript. The upstream region of the RNA heading toward the 5′ end is cleaved and dissociated from the EC, with no effect on the stability and activity of the EC. Most of the current data suggest that an 8- to 10-nt RNA⋅DNA hybrid is formed in the EC. Here, we show that an 8- to 10-nt RNA obtained by truncating the RNase-generated products further with either GreB or pyrophosphate is sufficient for the high stability and activity of the EC. This result suggests that the transcript–RNAP interaction that is required for holding the EC together can be limited to the RNA region involved in the 8- to 10-nt RNA⋅DNA hybrid.

Bibliography

Komissarova, N., & Kashlev, M. (1998). Functional topography of nascent RNA in elongation intermediates of RNA polymerase. Proceedings of the National Academy of Sciences, 95(25), 14699â14704.

Authors 2

Natalia Komissarova (first)
Mikhail Kashlev (additional)

References 33 Referenced 97

10.1146/annurev.biochem.66.1.117
T D Yager, P von Hippel Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology, eds F C Neidhardt, J L Ingraham, K B Low, B Magasanik, M Schaechter, H E Umbarger (Am. Soc. Microbiol., Washington, DC), pp. 1241–1275 (1987). / Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology by Yager T D (1987)
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C L Chan, R Landick Transcription: Mechanisms and Regulation, eds R C Conaway, J W Conaway (Raven, New York), pp. 297–321 (1994). / Transcription: Mechanisms and Regulation by Chan C L (1994)
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10.1016/0014-5793(82)80323-2

Dates

Type	When
Created	23 years, 1 month ago (July 26, 2002, 10:40 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 4:59 p.m.)
Indexed	1 month, 1 week ago (July 25, 2025, 6:51 a.m.)
Issued	26 years, 8 months ago (Dec. 8, 1998)
Published	26 years, 8 months ago (Dec. 8, 1998)
Published Online	26 years, 8 months ago (Dec. 8, 1998)
Published Print	26 years, 8 months ago (Dec. 8, 1998)

Funders 0

None

BibTeX

@article{Komissarova_1998, title={Functional topography of nascent RNA in elongation intermediates of RNA polymerase}, volume={95}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.95.25.14699}, DOI={10.1073/pnas.95.25.14699}, number={25}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Komissarova, Natalia and Kashlev, Mikhail}, year={1998}, month=dec, pages={14699–14704} }

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