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

J. Biol. Chem., Vol. 277, Issue 19, 16758-16767, May 10, 2002

This Article Full Text Full Text (PDF) All Versions of this Article: 277/19/16758    most recent M200707200v1 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 Serre, M.-C. Articles by Duguet, M. Search for Related Content PubMed PubMed Citation Articles by Serre, M.-C. Articles by Duguet, M.

Cleavage Properties of an Archaeal Site-specific Recombinase, the SSV1 Integrase^*

Marie-Claude Serre^§¶, Claire Letzelter^§, Jean-Renaud Garel^**, and Michel Duguet^§

From the  Laboratoire d'Enzymologie et Biochimie Structurales, CNRS Bat. 34, avenue de la Terrasse, 91198 Gif-sur-Yvette Cedex, France and the ^§ Laboratoire d'Enzymologie des Acides Nucléiques, Institut de Génétique et Microbiologie bât 400, Université Paris Sud, 91405 Orsay Cedex, France

SSV1 is a virus infecting the extremely thermophilic archaeon Sulfolobus shibatae. The viral-encoded integrase is responsible for site-specific integration of SSV1 into its host genome. The recombinant enzyme was expressed in Escherichia coli, purified to homogeneity, and its biochemical properties investigated in vitro. We show that the SSV1 integrase belongs to the tyrosine recombinases family and that Tyr³¹⁴ is involved in the formation of a 3'-phosphotyrosine intermediate. The integrase cleaves both strands of a synthetic substrate in a temperature-dependent reaction, the cleavage efficiency increasing with temperature. A discontinuity was observed in the Arrhenius plot above 50 °C, suggesting that a conformational transition may occur in the integrase at this temperature. Analysis of cleavage time course suggested that noncovalent binding of the integrase to its substrate is rate-limiting in the cleavage reaction. The cleavage positions were localized on each side of the anticodon loop of the tRNA gene where SSV1 integration takes place. Finally, the SSV1 integrase is able to cut substrates harboring mismatches in the binding site. For the cleavage step, the chemical nature of the base in position 1 of cleavage seems to be more important than its pairing to the opposite strand.

This article has been cited by other articles:

				X. Peng Evidence for the horizontal transfer of an integrase gene from a fusellovirus to a pRN-like plasmid within a single strain of Sulfolobus and the implications for plasmid survival Microbiology, February 1, 2008; 154(2): 383 - 391. [Abstract] [Full Text] [PDF]

				G. Erauso, K. M. Stedman, H. J. G. van de Werken, W. Zillig, and J. van der Oost Two novel conjugative plasmids from a single strain of Sulfolobus Microbiology, July 1, 2006; 152(7): 1951 - 1968. [Abstract] [Full Text] [PDF]

				C. Letzelter, M. Duguet, and M.-C. Serre Mutational Analysis of the Archaeal Tyrosine Recombinase SSV1 Integrase Suggests a Mechanism of DNA Cleavage in trans J. Biol. Chem., July 9, 2004; 279(28): 28936 - 28944. [Abstract] [Full Text] [PDF]

				B. Wiedenheft, K. Stedman, F. Roberto, D. Willits, A.-K. Gleske, L. Zoeller, J. Snyder, T. Douglas, and M. Young Comparative Genomic Analysis of Hyperthermophilic Archaeal Fuselloviridae Viruses J. Virol., February 15, 2004; 78(4): 1954 - 1961. [Abstract] [Full Text] [PDF]