title: The Bloom's syndrome gene product interacts with topoisomerase III.
abstract: Bloom's syndrome is a rare genetic disorder associated with loss of genomic integrity and a large increase in the incidence of many types of cancer at an early age. The Bloom's syndrome gene product, BLM, belongs to the RecQ family of DNA helicases, which also includes the human Werner's and Rothmund-Thomson syndrome gene products and the Sgs1 protein of Saccharomyces cerevisiae. This family shows strong evolutionary conservation of protein structure and function. Previous studies have shown that Sgs1p interacts both physically and genetically with topoisomerase III. Here, we have investigated whether this interaction has been conserved in human cells. We show that BLM and hTOPO IIIalpha, one of two human topoisomerase III homologues, co-localize in the nucleus of human cells and can be co-immunoprecipitated from human cell extracts. Moreover, the purified BLM and hTOPO IIIalpha proteins are able to bind specifically to each other in vitro, indicating that the interaction is direct. We have mapped two independent domains on BLM that are important for mediating the interaction with hTOPO IIIalpha. Furthermore, through characterizing a genetic interaction between BLM and TOP3 in S. cerevisiae, we have identified a functional role for the hTOPO IIIalpha interaction domains in BLM.
authors.affiliation: Imperial Cancer Research Fund Laboratories, Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital, Oxford OX3 9DS, United Kingdom.
authors.forename: L
authors.initials: L
authors.lastname: Wu
authors.forename: S L
authors.initials: SL
authors.lastname: Davies
authors.forename: P S
authors.initials: PS
authors.lastname: North
authors.forename: H
authors.initials: H
authors.lastname: Goulaouic
authors.forename: J F
authors.initials: JF
authors.lastname: Riou
authors.forename: H
authors.initials: H
authors.lastname: Turley
authors.forename: K C
authors.initials: KC
authors.lastname: Gatter
authors.forename: I D
authors.initials: ID
authors.lastname: Hickson
mesh_terms: D000251:Adenosine Triphosphatases / Q000276:immunology / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D001816:Bloom Syndrome / Q000378:metabolism*; D002461:Cell Line, Transformed; D004265:DNA Helicases / Q000276:immunology / Q000378:metabolism*; D017931:DNA Primers; D004264:DNA Topoisomerases, Type I / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D011485:Protein Binding; D053484:RecQ Helicases; D012441:Saccharomyces cerevisiae / Q000201:enzymology
chemical_list: D000911:Antibodies, Monoclonal; D017931:DNA Primers; D000251:Adenosine Triphosphatases; C097151:Bloom syndrome protein; D004265:DNA Helicases; D053484:RecQ Helicases; D004264:DNA Topoisomerases, Type I