Crystal structure of Sulfolobus solfataricus topoisomerase III reveals a novel carboxyl-terminal zinc finger domain essential for decatenation activity

Abstract

DNA topoisomerases are essential enzymes for a variety of cellular processes involved in DNA transactions. Mechanistic insights into type IA DNA topoisomerases have come principally from studies on bacterial and eukaryotic enzymes. A structural understanding of type IA topoisomerases in Archaea is lacking. Here, we present a 2.1-angstrom crystal structure of full-length Sulfolobus solfataricus topoisomerase III (Sso topo III), an archaeal member of type IA topoisomerases. The structure shows that Sso topo III adopts a characteristic torus-like architecture consisting of a four-domain core region and a novel carboxyl-terminal zinc finger domain (domain V). Structure-based mutation analyses reveal that a novel zinc-binding motif in domain V is essential for the DNA decatenation activity of Sso topo III. Our data indicate that Sso topo III represents a subclass of Type IA topoisomerases capable of resolving DNA catenates using a domain V-dependent mechanism. IMPORTANCEType IA topoisomerases are omnipresent in all cellular life forms and serve pivotal roles in cellular processes involved in DNA transactions. While considerable insights have been gained into Type IA topoisomerases from bacteria and eukaryotes, a structural understanding of type IA topoisomerases in Archaea remains elusive. we first determined the crystal structure of full-length Sulfolobus solfataricus topoisomerase III (Sso topo III), an archaeal member of type IA topoisomerases. Our structure provides the first molecular view of this archaeal topoisomerase, which removes negative supercoils and decatenates DNA catenane. Our findings manifest that Sso topo III may serve as an alternative prototype of type IA topoisomerases, whose decatenation mechanism differs from that of known bacterial and eukaryotic topoisomerases III such as Escherichia coli topoisomerase III (EcTOP3).