Alpha 2.0
Login Register
» Articles » PMID: 2987879

Influence of Monovalent Cations on the Activity of T4 DNA Ligase in the Presence of Polyethylene Glycol

Overview
Journal Nucleic Acids Res
Publisher Oxford University Press
Specialty Biochemistry
Date 1985 May 10
PMID 2987879
Citations 15
Authors
K Hayashi
M Nakazawa
Y Ishizaki
A OBAYASHI
Affiliations
Soon will be listed here.
Abstract

Monovalent cations such as Na+ and K+ inhibit the activity of T4 DNA ligase. However, the extent of inhibition varies with the terminal sequence of the duplex DNA used as substrate; in many cases, ligation of DNA is completely inhibited at 200 mM. The activity of the ligase is stimulated by raising the concentration of polyethylene glycol 6000 from 0 to 15% (w/v) when NaC1 and KC1 were both absent. Ligation was reduced as the concentration of NaC1 or KC1 was raised in a mixture containing 5 or 15% PEG 6000. With 10% PEG 6000, both cohesive- and blunt-end ligation of this ligase increased at high concentrations of salt (150-200 mM NaC1, or 200-250 mM KC1). Further, with 10% PEG 6000, inter- and intramolecular ligation occurred at low salt concentrations (0-100 mM NaC1, or 0-150 mM KC1); only linear oligomers were formed by intermolecular ligation at the high concentrations.

Citing Articles

Investigation of the Basic Steps in the Chromosome Conformation Capture Procedure.

Bylino O, Ibragimov A, Pravednikova A, Shidlovskii Y Front Genet. 2021; 12:733937.

PMID: 34616432 PMC: 8488379. DOI: 10.3389/fgene.2021.733937.

A single-molecule sequencing assay for the comprehensive profiling of T4 DNA ligase fidelity and bias during DNA end-joining.

Potapov V, Ong J, Langhorst B, Bilotti K, Cahoon D, Canton B Nucleic Acids Res. 2018; 46(13):e79.

PMID: 29741723 PMC: 6061786. DOI: 10.1093/nar/gky303.

Efficient DNA ligation by selective heating of DNA ligase with a radio frequency alternating magnetic field.

Suzuki M, Hayashi H, Mizuki T, Maekawa T, Morimoto H Biochem Biophys Rep. 2017; 8:360-364.

PMID: 28955977 PMC: 5614472. DOI: 10.1016/j.bbrep.2016.10.006.

Kinetics of T3-DNA Ligase-Catalyzed Phosphodiester Bond Formation Measured Using the α-Hemolysin Nanopore.

Tan C, Riedl J, Fleming A, Burrows C, White H ACS Nano. 2016; 10(12):11127-11135.

PMID: 28024377 PMC: 5302010. DOI: 10.1021/acsnano.6b05995.

Human DNA ligase III bridges two DNA ends to promote specific intermolecular DNA end joining.

Kukshal V, Kim I, Hura G, Tomkinson A, Tainer J, Ellenberger T Nucleic Acids Res. 2015; 43(14):7021-31.

PMID: 26130724 PMC: 4538836. DOI: 10.1093/nar/gkv652.

References
1.
Tsurimoto T, Matsubara K . Replication of lambda dv plasmid in vitro promoted by purified lambda O and P proteins. Proc Natl Acad Sci U S A. 1982; 79(24):7639-43. PMC: 347403. DOI: 10.1073/pnas.79.24.7639. View
2.
Zimmerman S, Pheiffer B . Macromolecular crowding allows blunt-end ligation by DNA ligases from rat liver or Escherichia coli. Proc Natl Acad Sci U S A. 1983; 80(19):5852-6. PMC: 390173. DOI: 10.1073/pnas.80.19.5852. View
3.
Weiss B, Richardson C . Enzymatic breakage and joining of deoxyribonucleic acid, I. Repair of single-strand breaks in DNA by an enzyme system from Escherichia coli infected with T4 bacteriophage. Proc Natl Acad Sci U S A. 1967; 57(4):1021-8. PMC: 224649. DOI: 10.1073/pnas.57.4.1021. View
4.
Weiss B, Jacquemin-Sablon A, Live T, Fareed G, Richardson C . Enzymatic breakage and joining of deoxyribonucleic acid. VI. Further purification and properties of polynucleotide ligase from Escherichia coli infected with bacteriophage T4. J Biol Chem. 1968; 243(17):4543-55. View
5.
Weiss B, Thompson A, Richardson C . Ezymatic breakage and joining of deoxyribonucleic acid. VII. Properties of the enzyme-adenylate intermediate in the polynucleotide ligase reaction. J Biol Chem. 1968; 243(17):4556-63. View