PMID: 9182992May 1, 1997Paper

Identification of amino acids of endonuclease VII essential for binding and cleavage of cruciform DNA

European Journal of Biochemistry
S GolzB Kemper

Abstract

Endonuclease VII is a Holliday-structure-resolving enzyme of bacteriophage T4. The active protein is a homodimer with 157 amino acids/monomer. An amber mutation (amE727 in codon 151) inactivates the nuclease completely, indicating the importance of the seven C-terminal amino acids for nucleolytic activity. The influence of these amino acids on cruciform-DNA binding and cleavage was investigated through functional analysis of C-terminal-truncated proteins derived from deletion constructs. It was found that the three C-terminal amino acids are not necessary for binding and cleavage. A transition from active to inactive protein occurs gradually with truncations of the next four amino acids. Reduction of DNA-binding ability, as measured by electrophoretic mobility shift assays, was determined to be the primary defect in the cleavage-deficient proteins. This was further concluded by the finding that EVII-(1-150)-peptide(amber), a protein with fairly low affinity to cruciform DNA, contributes cleavage activity to reactions of wild-type EVII with cruciform DNA. [Asp62]EVII-(1-156)-peptide lacking one C-terminal amino acid, contains a point mutation in codon 62 that eliminates the nucleolytic activity of the protein while retaining its...Continue Reading

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Citations

Jan 1, 1997·Biopolymers·D Suck
Jul 26, 2002·Journal of Molecular Biology·Rainer P Birkenbihl, Börries Kemper
Mar 20, 1999·Genetic Analysis : Biomolecular Engineering·G R Taylor, J Deeble
Jun 5, 2001·Nature Reviews. Molecular Cell Biology·D M Lilley, M F White
Mar 11, 2003·Microbiology and Molecular Biology Reviews : MMBR·Eric S MillerWolfgang Rüger
May 8, 2007·Virology·Gipsi Lima-MendezRaphaël Leplae
Sep 18, 2007·Nature·Christian BiertümpfelDietrich Suck
Nov 21, 1998·The Journal of Biological Chemistry·C KupferB Kemper
May 18, 2005·The Journal of Biological Chemistry·Mahua GhoshLars C Pedersen
Jun 18, 2017·Nature Communications·Marek SebestaDragana Ahel
Jul 18, 2001·The Journal of Biological Chemistry·Z ZhangY Chen
Dec 30, 2017·Journal of the American Chemical Society·Hisae Tateishi-KarimataNaoki Sugimoto

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