DOI: 10.1073/pnas.94.22.11869. Extensive purification of a putative RNA polymerase I holoenzyme from plants that accurately initiates rRNA gene transcription <i>in vitro</i>

Extensive purification of a putative RNA polymerase I holoenzyme from plants that accurately initiates rRNA gene transcription in vitro

10.1073/pnas.94.22.11869

Crossref journal-article

Proceedings of the National Academy of Sciences

Proceedings of the National Academy of Sciences (341)

Abstract

RNA polymerase I (pol I) is a nuclear enzyme whose function is to transcribe the duplicated genes encoding the precursor of the three largest ribosomal RNAs. We report a cell-free system from broccoli ( Brassica oleracea ) inflorescence that supports promoter-dependent RNA pol I transcription in vitro . The transcription system was purified extensively by DEAE-Sepharose, Biorex 70, Sephacryl S300, and Mono Q chromatography. Activities required for pre-rRNA transcription copurified with the polymerase on all four columns, suggesting their association as a complex. Purified fractions programmed transcription initiation from the in vivo start site and utilized the same core promoter sequences required in vivo . The complex was not dissociated in 800 mM KCl and had a molecular mass of nearly 2 MDa based on gel filtration chromatography. The most highly purified fractions contain ≈30 polypeptides, two of which were identified immunologically as RNA polymerase subunits. These data suggest that the occurrence of a holoenzyme complex is probably not unique to the pol II system but may be a general feature of eukaryotic nuclear polymerases.

Bibliography

Saez-Vasquez, J., & Pikaard, C. S. (1997). Extensive purification of a putative RNA polymerase I holoenzyme from plants that accurately initiates rRNA gene transcription inâvitro. Proceedings of the National Academy of Sciences, 94(22), 11869â11874.

Authors 2

Julio Saez-Vasquez (first)
Craig S. Pikaard (additional)

References 49 Referenced 51

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Dates

Type	When
Created	23 years ago (July 26, 2002, 10:40 a.m.)
Deposited	3 years, 4 months ago (April 13, 2022, 3:21 p.m.)
Indexed	2 weeks, 4 days ago (Aug. 5, 2025, 8:15 a.m.)
Issued	27 years, 9 months ago (Oct. 28, 1997)
Published	27 years, 9 months ago (Oct. 28, 1997)
Published Online	27 years, 9 months ago (Oct. 28, 1997)
Published Print	27 years, 9 months ago (Oct. 28, 1997)

Funders 0

None

BibTeX

@article{Saez_Vasquez_1997, title={Extensive purification of a putative RNA polymerase I holoenzyme from plants that accurately initiates rRNA gene transcription in vitro}, volume={94}, ISSN={1091-6490}, url={http://dx.doi.org/10.1073/pnas.94.22.11869}, DOI={10.1073/pnas.94.22.11869}, number={22}, journal={Proceedings of the National Academy of Sciences}, publisher={Proceedings of the National Academy of Sciences}, author={Saez-Vasquez, Julio and Pikaard, Craig S.}, year={1997}, month=oct, pages={11869–11874} }

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