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J Biol Chem, Vol. 275, Issue 13, 9636-9644, March 31, 2000
,,,
From the 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 III Imperial Cancer Research Fund Laboratories,
Institute of Molecular Medicine, University of Oxford, John Radcliffe
Hospital, Oxford OX3 9DS, United Kingdom,
§ Rhône-Poulenc Rorer, Centre de Recherche de
Vitry-Alfortville, Vitry sur Seine Cedex, 94403 France, and the
¶ Department of Cellular Science, University of Oxford, John
Radcliffe Hospital, Oxford OX3 9DU, United Kingdom
, 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 III 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 III.
Furthermore, through characterizing a genetic interaction between
BLM and TOP3 in S. cerevisiae, we
have identified a functional role for the hTOPO III interaction
domains in BLM.
To whom correspondence should be addressed. Tel.:
44-1865-222417; Fax: 44-1865-222431; E-mail:
hickson@icrf.icnet.uk.
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