Kinetic Analysis of the Exonuclease Activity of the Bacteriophage T4 Mre11-Rad50 Complex.

Authors :: Teklemariam TA
Rivera OD
Nelson SW
Source :: Methods in enzymology [Methods Enzymol] 2018; Vol. 600, pp. 135-156. Date of Electronic Publication: 2018 Feb 03.
Publication Year :: 2018
Abstract: Bacteriophage T4 encodes orthologs of the proteins Rad50 (gp46) and Mre11 (gp47), which form a heterotetrameric complex (MR) that is responsible for host genome degradation and the processing of DNA ends for recombination-dependent DNA repair. In this chapter, we describe the ensemble methods currently employed by our laboratory to characterize the exonuclease activity of the T4 MR complex. DNA exonucleases play a vital role in maintaining the integrity of DNA through their participation in DNA repair pathways and as proofreaders for DNA polymerases. Methods for quantifying the general features of the exonuclease, and for determining steady-state kinetic parameters (K <subscript>m</subscript> , k <subscript>cat</subscript> ), the polarity of exonuclease activity, and processivity are presented. These methods should be applicable to all DNA exonucleases, and to some extent endonucleases.<br /> (© 2018 Elsevier Inc. All rights reserved.)

Subjects :: Bacteriophage T4 metabolism
Chromatography, Thin Layer instrumentation
Chromatography, Thin Layer methods
DNA, Single-Stranded chemistry
DNA, Single-Stranded genetics
Electrophoresis, Agar Gel instrumentation
Electrophoresis, Agar Gel methods
Enzyme Assays instrumentation
Kinetics
Staining and Labeling instrumentation
Staining and Labeling methods
Bacteriophage T4 genetics
DNA, Single-Stranded metabolism
Enzyme Assays methods
Recombinational DNA Repair
Viral Proteins metabolism

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