Synergic and opposing activities of thermophilic RecQ-like helicase and topoisomerase 3 proteins in Holliday junction processing and replication fork stabilization.

Abstract

RecQ family helicases and topoisomerase 3 enzymes form evolutionary conserved complexes that play essential functions in DNA replication, recombination, and repair, and in vitro, show coordinate activities on model recombination and replication intermediates. Malfunctioning of these complexes in humans is associated with genomic instability and cancer-prone syndromes. Although both RecQ-like and topoisomerase 3 enzymes are present in archaea, only a few of them have been studied, and no information about their functional interaction is available. We tested the combined activities of the RecQ-like helicase, Hel112, and the topoisomerase 3, SsTop3, from the thermophilic archaeon Sulfolobus solfataricus. Hel112 showed coordinate DNA unwinding and annealing activities, a feature shared by eukaryotic RecQ homologs, which resulted in processing of synthetic Holliday junctions and stabilization of model replication forks. SsTop3 catalyzed DNA relaxation and annealing. When assayed in combination, SsTop3 inhibited the Hel112 helicase activity on Holliday junctions and stimulated formation and stabilization of such structures. In contrast, Hel112 did not affect the SsTop3 DNA relaxation activity. RecQ-topoisomerase 3 complexes show structural similarity with the thermophile-specific enzyme reverse gyrase, which catalyzes positive supercoiling of DNA and was suggested to play a role in genome stability at high temperature. Despite such similarity and the high temperature of reaction, the SsTop3-Hel112 complex does not induce positive supercoiling and is thus likely to play different roles. We propose that the interplay between Hel112 and SsTop3 might regulate the equilibrium between recombination and anti-recombination activities at replication forks.

• Authors: Anna Valenti, Mariarita De Felice, Giuseppe Perugino, Anna Bizard, Marc Nadal, Mosè Rossi, Maria Ciaramella
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 287 Issue 36 Pg. 30282-95 (Aug 31 2012) ISSN: 1083-351X [Electronic] United States
• PMID: 22722926 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Archaeal Proteins
• DNA, Archaeal
• DNA, Cruciform
• RecQ Helicases
• DNA Topoisomerases, Type I

Topics

• Archaeal Proteins (genetics, metabolism)
• DNA Replication (physiology)
• DNA Topoisomerases, Type I (genetics, metabolism)
• DNA, Archaeal (biosynthesis, genetics)
• DNA, Cruciform (genetics, metabolism)
• RecQ Helicases (genetics, metabolism)
• Sulfolobus solfataricus (enzymology, genetics)