Abstract
Yeast mitochondrial RNA polymerase can bind specifically to promoter-containing DNA fragments in vitro as detected by DNAse I or methidiumpropyl-EDTA. Fe(II) protection assays and gel retardation experiments. Retardation of RNA polymerase-DNA complexes was most pronounced when the promoter was located in the middle of a DNA fragment and was diminished when RNA polymerase was bound near one of the ends. This indicates that upon RNA polymerase-binding the DNA undergoes a conformational change which is most likely a bend. The degree of introduced bending correlated with the efficiency of transcription and promoter-binding in a series of promoter mutants, suggesting that bending is a functional event during promoter utilisation.
MeSH Terms
DNA Mutational Analysis
DNA, Mitochondrial/metabolism,ultrastructure
DNA-Binding Proteins/metabolism
DNA-Directed RNA Polymerases/metabolism
Nucleic Acid Conformation
Promoter Regions, Genetic
Protein Binding
Saccharomyces cerevisiae/genetics
Structure-Activity Relationship
Transcription, Genetic
Chemicals
DNA, Mitochondrial
DNA-Binding Proteins
DNA-Directed RNA Polymerases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Schinkel A H
Laboratory of Biochemistry, University of Amsterdam, The Netherlands.
Groot Koerkamp M J
Teunissen A W
Tabak H F
References (19)
19 references, click to expand
-
Mechanism and control of transcription initiation in prokaryotes.
Annu Rev Biochem. 1985;54:171-204
PMID: 3896120
-
In vitro characterization of the yeast mitochondrial promoter using single-base substitution mutants.
J Biol Chem. 1987 Oct 5;262(28):13690-6
PMID: 3308881
-
Intermediates in transcription initiation from the E. coli lac UV5 promoter.
Cell. 1985 Dec;43(2 Pt 1):449-59
PMID: 2416465
-
DNA bending at adenine . thymine tracts.
Nature. 1986 Apr 10-16;320(6062):501-6
PMID: 3960133
-
Design of sequence-specific DNA-binding molecules.
Science. 1986 Apr 25;232(4749):464-71
PMID: 2421408
-
Interactions of the RNA polymerase of bacteriophage T7 with its promoter during binding and initiation of transcription.
Proc Natl Acad Sci U S A. 1986 Jun;83(11):3614-8
PMID: 3459146
-
Characterization of the promoter of the large ribosomal RNA gene in yeast mitochondria and separation of mitochondrial RNA polymerase into two different functional components.
EMBO J. 1986 May;5(5):1041-7
PMID: 3522220
-
Yeast mitochondrial RNA polymerase. Purification and properties of the catalytic subunit.
J Biol Chem. 1986 Aug 5;261(22):10340-7
PMID: 3525543
-
Genetics of mitochondrial biogenesis.
Annu Rev Biochem. 1986;55:249-85
PMID: 2427014
-
Bending of promoter DNA on binding of heat shock transcription factor.
Nature. 1986 Oct 2-8;323(6087):459-61
PMID: 3020432
-
The DNA binding domain and bending angle of E. coli CAP protein.
Cell. 1986 Dec 26;47(6):995-1005
PMID: 3536129
-
Functional dissection of Escherichia coli promoters: information in the transcribed region is involved in late steps of the overall process.
EMBO J. 1986 Nov;5(11):2995-3000
PMID: 3539590
-
The gamma delta resolvase induces an unusual DNA structure at the recombinational crossover point.
Cell. 1987 Apr 10;49(1):103-10
PMID: 3030563
-
The integration host factor of Escherichia coli binds to bent DNA at the origin of replication of the plasmid pSC101.
Cell. 1987 Jun 5;49(5):709-17
PMID: 3555843
-
Unusual properties of promoter-up mutations in the Escherichia coli galactose operon and evidence suggesting RNA polymerase-induced DNA bending.
EMBO J. 1987 Feb;6(2):507-13
PMID: 3034593
-
Effect of point mutations on in vitro transcription from the promoter for the large ribosomal RNA gene of yeast mitochondria.
Nucleic Acids Res. 1987 Jul 24;15(14):5597-612
PMID: 3302943
-
Specificity factor of yeast mitochondrial RNA polymerase. Purification and interaction with core RNA polymerase.
J Biol Chem. 1987 Sep 15;262(26):12785-91
PMID: 2442167
-
Yeast mitochondrial RNA polymerase is homologous to those encoded by bacteriophages T3 and T7.
Cell. 1987 Oct 9;51(1):89-99
PMID: 3308116
-
A multicomponent mitochondrial RNA polymerase from Saccharomyces cerevisiae.
J Biol Chem. 1985 Nov 15;260(26):14214-23
PMID: 3902826