On the influence of thymidine analogues on the activity of phage fd promoters in vitro
Open Access
- 1 January 1980
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 8 (24) , 6143-6162
- https://doi.org/10.1093/nar/8.24.6143
Abstract
RF I DNA of phage fd containing 5-bromo-deoxyuridine (br5Ud) or deoxyuridine (Ud) instead of deoxythymidine (Td) in the codogenic strand was synthesized in vitro. The modified genomes could be cleaved by restriction endonuclease Hpa II. Although the recognition site of Hpa II is CCGG, the cleavage rate was significantly reduced with Ud-containing DNA. Both base substitutions altered the mobilities of several DNA fragments under the conditions of polyacrylamide gel electrophoresis. The fragments containing binding sites for RNA polymerase were assayed for the rates of stable complex formation. The substitution of Td for both, Ud and br 5Ud, strongly influenced this parameter. Thus the methyl group of Td has to be regarded as one of the sites in DNA which determine the rate of stable RNA polymerase binding and thereby possibly mediate promoter activity in vitro (24, 25, 26). In most cases the rate of complex formation was decreased by Ud, but increased by br 5Ud.Keywords
This publication has 41 references indexed in Scilit:
- Studies on bacteriophage fd DNA: II. Localization of RNA initiation sites on the cleavage map of the fd genomeJournal of Molecular Biology, 1975
- Mapping and characterization of promoters in bacteriophages fd, f1 and M13Journal of Molecular Biology, 1975
- Synthesis and Nucleolytic Degradation of Uracil-containing Deoxyribonucleic Acid by Escherichia coli Deoxyribonucleic Acid Polymerase IJournal of Biological Chemistry, 1973
- Lac Operator Analogues: Bromodeoxyuridine Substitution in the lac Operator Affects the Rate of Dissociation of the lac RepressorProceedings of the National Academy of Sciences, 1972
- Selective Elimination of the Exonuclease Activity of the Deoxyribonucleic Acid Polymerase from Escherichia coli B by Limited ProteolysisProceedings of the National Academy of Sciences, 1970
- An active fragment of DNA polymerase produced by proteolytic cleavageBiochemical and Biophysical Research Communications, 1969
- Starting nucleotide sequences of RNA synthesized on the replicative form DNA of coliphage fdJournal of Molecular Biology, 1969
- Studies on the binding of RNA polymerase to polynucleotidesJournal of Molecular Biology, 1966
- Enzymic synthesis of RNA from T7 DNAJournal of Molecular Biology, 1966
- Preparation of double-stranded DNA (replicative form) of bacteriophage ΦX174: A simplified methodBiochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis, 1966