Three-dimensional structure of Escherichia coli RNA polymerase holoenzyme determined by electron crystallography - PubMed (original) (raw)
. 1989 Aug 31;340(6236):730-2.
doi: 10.1038/340730a0.
Affiliations
- PMID: 2671751
- DOI: 10.1038/340730a0
Three-dimensional structure of Escherichia coli RNA polymerase holoenzyme determined by electron crystallography
S A Darst et al. Nature. 1989.
Abstract
During transcription in E. coli, the DNA-dependent RNA polymerase locates specific promoter sequences in the DNA template, melts a small region containing the transcription start site, initiates RNA synthesis, processively elongates the transcript, and finally terminates and releases the RNA product. Each step is regulated by interactions between the polymerase, the DNA, the nascent RNA, and a variety of regulatory proteins and ligands. The E. coli enzyme contains a catalytic core of two alpha-subunits, one beta- and one beta'-subunit, with relative molecular masses (Mr) of 36,512, 150,619 and 155,162, respectively. The holoenzyme has an additional regulatory subunit, normally sigma, of Mr 70,236. Preparations may also contain the omega-subunit (Mr approximately 10,000), which can be removed without affecting any known properties of the enzyme. Because the amino-acid sequences of the beta- and beta'-subunits are homologous to those of the largest subunits of the yeast, Drosophila and murine RNA polymerases, it seems likely that essential features of the three-dimensional structure and catalytic mechanism of RNA polymerase are also conserved across species. Crystals of RNA polymerase suitable for X-ray analysis have not yet been obtained, but two-dimensional crystals of E. coli RNA polymerase holoenzyme can be grown on positively charged lipid layers. Electron microscopy of these crystals in negative stain shows the enzyme in projection as an irregularly shaped complex approximately 100 x 100 x 160 A in size. We have now determined the three-dimensional structure by electron microscopy of negatively stained, two-dimensional crystals tilted at various angles to the incident electron beam. We find a structure in RNA polymerase similar to the active-site cleft of DNA polymerase I. In the light of functional similarities between these two enzymes, together with other evidence, this probably identifies the active-site region of RNA polymerase.
Similar articles
- Insights into Escherichia coli RNA polymerase structure from a combination of x-ray and electron crystallography.
Darst SA, Polyakov A, Richter C, Zhang G. Darst SA, et al. J Struct Biol. 1998 Dec 15;124(2-3):115-22. doi: 10.1006/jsbi.1998.4057. J Struct Biol. 1998. PMID: 10049799 - Interplay between the beta' clamp and the beta' jaw domains during DNA opening by the bacterial RNA polymerase at sigma54-dependent promoters.
Wigneshweraraj SR, Savalia D, Severinov K, Buck M. Wigneshweraraj SR, et al. J Mol Biol. 2006 Jun 23;359(5):1182-95. doi: 10.1016/j.jmb.2006.04.063. Epub 2006 May 15. J Mol Biol. 2006. PMID: 16725156 - Subunit of assembly of Escherichia coli RNA polymerase.
Ishihama A. Ishihama A. Adv Biophys. 1981;14:1-35. Adv Biophys. 1981. PMID: 7015808 Review. - Structural studies of Escherichia coli RNA polymerase.
Darst SA, Polyakov A, Richter C, Zhang G. Darst SA, et al. Cold Spring Harb Symp Quant Biol. 1998;63:269-76. doi: 10.1101/sqb.1998.63.269. Cold Spring Harb Symp Quant Biol. 1998. PMID: 10384291 Review. No abstract available.
Cited by
- RpoZ regulates 2,4-DAPG production and quorum sensing system in Pseudomonas fluorescens 2P24.
Wei Y, Dong B, Wu X, Zhao M, Wang D, Li N, Zhang Q, Zhang L, Zhou H. Wei Y, et al. Front Microbiol. 2023 May 12;14:1160913. doi: 10.3389/fmicb.2023.1160913. eCollection 2023. Front Microbiol. 2023. PMID: 37250031 Free PMC article. - Diverse and unified mechanisms of transcription initiation in bacteria.
Chen J, Boyaci H, Campbell EA. Chen J, et al. Nat Rev Microbiol. 2021 Feb;19(2):95-109. doi: 10.1038/s41579-020-00450-2. Epub 2020 Oct 29. Nat Rev Microbiol. 2021. PMID: 33122819 Free PMC article. Review. - Genome-wide mapping of Topoisomerase I activity sites reveal its role in chromosome segregation.
Rani P, Nagaraja V. Rani P, et al. Nucleic Acids Res. 2019 Feb 20;47(3):1416-1427. doi: 10.1093/nar/gky1271. Nucleic Acids Res. 2019. PMID: 30566665 Free PMC article. - An Introduction to the Structure and Function of the Catalytic Core Enzyme of Escherichia coli RNA Polymerase.
Sutherland C, Murakami KS. Sutherland C, et al. EcoSal Plus. 2018 Aug;8(1):10.1128/ecosalplus.ESP-0004-2018. doi: 10.1128/ecosalplus.ESP-0004-2018. EcoSal Plus. 2018. PMID: 30109846 Free PMC article. Review. - Using synthetic bacterial enhancers to reveal a looping-based mechanism for quenching-like repression.
Brunwasser-Meirom M, Pollak Y, Goldberg S, Levy L, Atar O, Amit R. Brunwasser-Meirom M, et al. Nat Commun. 2016 Feb 2;7:10407. doi: 10.1038/ncomms10407. Nat Commun. 2016. PMID: 26832446 Free PMC article.
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Molecular Biology Databases