Identification of a Human Homologue of theSchizosaccharomyces pombe rad17 + Checkpoint Gene

doi:10.1074/jbc.273.29.18340

Identification of a Human Homologue of theSchizosaccharomyces pombe rad17⁺ Checkpoint Gene*

From the ^‡Department of Experimental Molecular Biology and the ^¶Department of Applied Molecular Biology, Janssen Research Foundation, Turnhoutseweg 30, B-2340 Beerse, Belgium

Abstract

In the fission yeast Schizosaccharomyces pombe the rad17 ⁺ gene is required for both the DNA damage-dependent and the DNA replication-dependent cell cycle checkpoints. We have identified a human cDNA homologue of the S. pombe rad17 ⁺ checkpoint gene, designatedHrad17. Hrad17 has 49% identity to theS. pombe rad17 ⁺ sequence at the DNA level and 49% identity and 72% similarity at the amino acid level. Northern blot analysis indicates elevated levels of expression in testis and in cancer cell lines. Chromosomal localization by fluorescence in situ hybridization indicates that Hrad17 is located on chromosome 4q13.3–21.2. This region is subject to loss of heterozygosity in several human cancers. To begin to understand the protein-protein interactions of the human checkpoint machinery, we have used the yeast two-hybrid system to examine potential interactions between Hrad1, Hrad9, and Hrad17. We demonstrate a physical interaction between Hrad17 and Hrad1 but no interaction with Hrad9.

Footnotes

↵* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
↵§ Present address: Cardiovascular Metabolism and Musculoskeletal Research Dept., Zeneca Pharmaceuticals, Alderley Edge, Cheshire, UK.
↵‖ To whom correspondence should be addressed. Tel.: 32-14-602618 or 32-14-605734; Fax: 32-14-606111; E-mail:wluyten{at}janbe.jnj.com.
↵1 The abbreviations used are: EST, expressed sequence tag; RACE, rapid amplification of cDNA ends; PCR, polymerase chain reaction; ORF, open reading frame; FISH, fluorescencein situ hybridization; DAPI, 4′,6-diamidino-2-phenylindole; kb, kilobase pair(s).
- Received December 29, 1997.
- Revision received April 22, 1998.
The American Society for Biochemistry and Molecular Biology, Inc.