Functional interaction of reverse gyrase with single-strand binding protein of the archaeon Sulfolobus

Reverse gyrase is a unique hyperthermophile-specific DNA topoisomerase that induces positive supercoiling. It is a modular enzyme composed of a topoisomerase IA and a helicase domain, which cooperate in the ATP-dependent positive supercoiling reaction. Although its physiological function has not bee...

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Main Authors:	Napoli, Alessandra, Valenti, Anna, Salerno, Vincenzo, Nadal, Marc, Garnier, Florence, Rossi, Mosè, Ciaramella, Maria
Format:	Online
Language:	English
Published:	Oxford University Press 2005
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC548347/

id	pubmed-548347
recordtype	oai_dc
spelling	pubmed-5483472005-02-10 Functional interaction of reverse gyrase with single-strand binding protein of the archaeon Sulfolobus Napoli, Alessandra Valenti, Anna Salerno, Vincenzo Nadal, Marc Garnier, Florence Rossi, Mosè Ciaramella, Maria Article Reverse gyrase is a unique hyperthermophile-specific DNA topoisomerase that induces positive supercoiling. It is a modular enzyme composed of a topoisomerase IA and a helicase domain, which cooperate in the ATP-dependent positive supercoiling reaction. Although its physiological function has not been determined, it can be hypothesized that, like the topoisomerase–helicase complexes found in every organism, reverse gyrase might participate in different DNA transactions mediated by multiprotein complexes. Here, we show that reverse gyrase activity is stimulated by the single-strand binding protein (SSB) from the archaeon Sulfolobus solfataricus. Using a combination of in vitro assays we analysed each step of the complex reverse gyrase reaction. SSB stimulates all the steps of the reaction: binding to DNA, DNA cleavage, strand passage and ligation. By co-immunoprecipitation of cell extracts we show that reverse gyrase and SSB assemble a complex in the presence of DNA, but do not make stable protein–protein interactions. In addition, SSB stimulates reverse gyrase positive supercoiling activity on DNA templates associated with the chromatin protein Sul7d. Furthermore, SSB enhances binding and cleavage of UV-irradiated substrates by reverse gyrase. The results shown here suggest that these functional interactions may have biological relevance and that the interplay of different DNA binding proteins might modulate reverse gyrase activity in DNA metabolic pathways. Oxford University Press 2005 2005-01-26 /pmc/articles/PMC548347/ /pubmed/15673717 http://dx.doi.org/10.1093/nar/gki202 Text en © The Author 2005. Published by Oxford University Press. All rights reserved
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Napoli, Alessandra Valenti, Anna Salerno, Vincenzo Nadal, Marc Garnier, Florence Rossi, Mosè Ciaramella, Maria
spellingShingle	Napoli, Alessandra Valenti, Anna Salerno, Vincenzo Nadal, Marc Garnier, Florence Rossi, Mosè Ciaramella, Maria Functional interaction of reverse gyrase with single-strand binding protein of the archaeon Sulfolobus
author_facet	Napoli, Alessandra Valenti, Anna Salerno, Vincenzo Nadal, Marc Garnier, Florence Rossi, Mosè Ciaramella, Maria
author_sort	Napoli, Alessandra
title	Functional interaction of reverse gyrase with single-strand binding protein of the archaeon Sulfolobus
title_short	Functional interaction of reverse gyrase with single-strand binding protein of the archaeon Sulfolobus
title_full	Functional interaction of reverse gyrase with single-strand binding protein of the archaeon Sulfolobus
title_fullStr	Functional interaction of reverse gyrase with single-strand binding protein of the archaeon Sulfolobus
title_full_unstemmed	Functional interaction of reverse gyrase with single-strand binding protein of the archaeon Sulfolobus
title_sort	functional interaction of reverse gyrase with single-strand binding protein of the archaeon sulfolobus
description	Reverse gyrase is a unique hyperthermophile-specific DNA topoisomerase that induces positive supercoiling. It is a modular enzyme composed of a topoisomerase IA and a helicase domain, which cooperate in the ATP-dependent positive supercoiling reaction. Although its physiological function has not been determined, it can be hypothesized that, like the topoisomerase–helicase complexes found in every organism, reverse gyrase might participate in different DNA transactions mediated by multiprotein complexes. Here, we show that reverse gyrase activity is stimulated by the single-strand binding protein (SSB) from the archaeon Sulfolobus solfataricus. Using a combination of in vitro assays we analysed each step of the complex reverse gyrase reaction. SSB stimulates all the steps of the reaction: binding to DNA, DNA cleavage, strand passage and ligation. By co-immunoprecipitation of cell extracts we show that reverse gyrase and SSB assemble a complex in the presence of DNA, but do not make stable protein–protein interactions. In addition, SSB stimulates reverse gyrase positive supercoiling activity on DNA templates associated with the chromatin protein Sul7d. Furthermore, SSB enhances binding and cleavage of UV-irradiated substrates by reverse gyrase. The results shown here suggest that these functional interactions may have biological relevance and that the interplay of different DNA binding proteins might modulate reverse gyrase activity in DNA metabolic pathways.
publisher	Oxford University Press
publishDate	2005
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC548347/
_version_	1611370827806146560

Functional interaction of reverse gyrase with single-strand binding protein of the archaeon Sulfolobus

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