Assembly of functional Escherichia coli RNA polymerase containing beta subunit fragments.
- 9 May 1995
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 92 (10), 4591-4595
- https://doi.org/10.1073/pnas.92.10.4591
Abstract
The Escherichia coli rpoB gene, which codes for the 1342-residue beta subunit of RNA polymerase (RNAP), contains two dispensable regions centered around codons 300 and 1000. To test whether these regions demarcate domains of the RNAP beta subunit, fragments encoded by segments of rpoB flanking the dispensable regions were individually overexpressed and purified. We show that these beta-subunit polypeptide fragments, when added with purified recombinant beta', sigma, and alpha subunits of RNAP, reconstitute a functional enzyme in vitro. These results demonstrate that the beta subunit is composed of at least three distinct domains and open another avenue for in vitro studies of RNAP assembly and structure.Keywords
This publication has 49 references indexed in Scilit:
- Structural analysis of ternary complexes of Escherichia coli RNA polymerase: Deoxyribonuclease I footprinting of defined complexesJournal of Molecular Biology, 1992
- Three-dimensional structure of yeast RNA polymerase II at 16 Å resolutionCell, 1991
- Evolution of RNA polymerases and branching patterns of the three major groups of archaebacteriaJournal of Molecular Evolution, 1991
- Blocking of the Initiation-to-Elongation Transition by a Transdominant RNA Polymerase MutationScience, 1990
- Three-dimensional structure of Escherichia coli RNA polymerase holoenzyme determined by electron crystallographyNature, 1989
- Mapping and sequencing of mutations in the Escherichia colirpoB gene that lead to rifampicin resistanceJournal of Molecular Biology, 1988
- Spinach chloroplast rpoBC genes encode three subunits of the chloroplast RNA polymeraseJournal of Molecular Biology, 1988
- RNA polymerase II of DrosophilaJournal of Molecular Biology, 1987
- Studies on the functional topography of Escherichia coli RNA polymeraseEuropean Journal of Biochemistry, 1987
- Structure of the eukaryotic transcription apparatus: Features of the gene for the largest subunit of Drosophila RNA polymerase IICell, 1985