Consequences of molecular engineering enhanced DNA binding in a DNA repair enzyme

Courtney Nickell, Melissa A. Prince, Robert (Stephen) Lloyd

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Facilitated one-dimensional diffusion is a general mechanism utilized by several DNA-interactive proteins as they search for their target sites within large domains of nontarget DNA. T4 endonuclease V is a protein which scans DNA in a nonspecifically bound state and processively incises DNA at ultraviolet (UV)-induced pyrimidine dimer sites. An electrostatic contribution to this mechanism of target location has been established. Previous studies indicate that a decrease in the affinity of endonuclease V for nontarget DNA results in a decreased ability to scan DNA and a concomitant decrease in the ability to enhance UV survival in repair-deficient Escherichia coli. This study was designed to question the contrasting effect of an increase in the affinity of endonuclease V for nontarget DNA. With this as a goal, a gradient of increasingly basic amino acid content was created along a proposed endonuclease V-nontarget DNA interface. This incremental increase in positive charge correlated with the stepwise enhancement of nontarget DNA binding, yet inversely correlated with enhanced UV survival in repair-deficient E. coli. Further analysis suggests that the observed reduction in UV survival is consistent with the hypothesis that enhanced nontarget DNA affinity results in reduced pyrimidine dimer-specific recognition and/or binding. The net effect is a reduction in the efficiency of pyrimidine dimer incision.

Original languageEnglish (US)
Pages (from-to)4189-4198
Number of pages10
JournalBiochemistry
Volume31
Issue number17
StatePublished - 1992
Externally publishedYes

Fingerprint

DNA Repair Enzymes
DNA
Deoxyribonuclease (Pyrimidine Dimer)
Pyrimidine Dimers
Escherichia coli
Repair
Basic Amino Acids
Static Electricity
Electrostatics
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Consequences of molecular engineering enhanced DNA binding in a DNA repair enzyme. / Nickell, Courtney; Prince, Melissa A.; Lloyd, Robert (Stephen).

In: Biochemistry, Vol. 31, No. 17, 1992, p. 4189-4198.

Research output: Contribution to journalArticle

Nickell, Courtney ; Prince, Melissa A. ; Lloyd, Robert (Stephen). / Consequences of molecular engineering enhanced DNA binding in a DNA repair enzyme. In: Biochemistry. 1992 ; Vol. 31, No. 17. pp. 4189-4198.
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