HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 276, Issue 13, 10432-10436, March 30, 2001

This Article Full Text Full Text (PDF) All Versions of this Article: 276/13/10432    most recent M010138200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Yoshikawa, M. Articles by Shinagawa, H. Search for Related Content PubMed PubMed Citation Articles by Yoshikawa, M. Articles by Shinagawa, H.

Evidence that Phenylalanine 69 in Escherichia coli RuvC Resolvase Forms a Stacking Interaction during Binding and Destabilization of a Holliday Junction DNA Substrate^*

Manabu Yoshikawa, Hiroshi Iwasaki^§¶, and Hideo Shinagawa

From the  Department of Molecular Microbiology, Research Institute for Microbial Diseases, Osaka University, and ^§ PRESTO, Japan Science and Technology Corporation, Suita, Osaka, 565-0871, Japan

Escherichia coli RuvC resolvase is a specific endonuclease that recognizes and cleaves Holliday junctions formed during homologous recombination and recombinational repair. This study examines the phenotype of RuvC mutants with amino acid substitutions at phenylalanine 69 (F69L, F69Y, F69W, and F69A), a catalytically important residue that faces the catalytic center of the enzyme. F69Y, but not the other three mutants, almost fully complements the UV sensitivity of a ruvC strain and substantially resolves synthetic Holliday junctions in vitro. In the presence of 100 mM NaCl, RuvC F69A and F69L are defective in junction binding, but F69Y and F69W retain near wild-type binding activity during a gel shift binding assay. KMnO₄ was used to probe synthetic Holliday junction DNA in a complex with wild-type and mutant RuvC; F69A and F69L did not induce disruption of base pairing at the crossover to the same extent as wild-type RuvC. Thus, the aromatic ring of Phe-69 is involved in DNA binding, probably via a stacking interaction with a nucleotide base, and this interaction may induce a structural change in junction DNA that is required to form a catalytically competent complex.

This article has been cited by other articles:

				K. V. Kepple, J. L. Boldt, and A. M. Segall Holliday junction-binding peptides inhibit distinct junction-processing enzymes PNAS, May 10, 2005; 102(19): 6867 - 6872. [Abstract] [Full Text] [PDF]

				E. Matsui, J. Abe, H. Yokoyama, and I. Matsui Aromatic Residues Located Close to the Active Center Are Essential for the Catalytic Reaction of Flap Endonuclease-1 from Hyperthermophilic Archaeon Pyrococcus horikoshii J. Biol. Chem., April 16, 2004; 279(16): 16687 - 16696. [Abstract] [Full Text] [PDF]

				M. Iwase, Y. Satta, Y. Hirai, H. Hirai, H. Imai, and N. Takahata From the Cover: The amelogenin loci span an ancient pseudoautosomal boundary in diverse mammalian species PNAS, April 29, 2003; 100(9): 5258 - 5263. [Abstract] [Full Text] [PDF]

				M. F. Loughlin, F. M. Barnard, D. Jenkins, G. J. Sharples, and P. J. Jenks Helicobacter pylori Mutants Defective in RuvC Holliday Junction Resolvase Display Reduced Macrophage Survival and Spontaneous Clearance from the Murine Gastric Mucosa Infect. Immun., April 1, 2003; 71(4): 2022 - 2031. [Abstract] [Full Text]

				T. Nishino, K. Komori, Y. Ishino, and K. Morikawa Dissection of the Regional Roles of the Archaeal Holliday Junction Resolvase Hjc by Structural and Mutational Analyses J. Biol. Chem., September 14, 2001; 276(38): 35735 - 35740. [Abstract] [Full Text] [PDF]