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J Biol Chem, Vol. 273, Issue 29, 18332-18339, July 17, 1998
From the In the fission yeast Schizosaccharomyces
pombe the rad1+ gene is required for both
the DNA damage-dependent and the DNA
replication-dependent cell cycle checkpoints. We have
identified a human homologue of the S. pombe
rad1+ gene, designated Hrad1, as well
as a mouse homologue: Mrad1. Two Hrad1
alternative splice variants with different open reading frames have
been identified; one codes for a long form, Hrad1A, and the other
encodes a short form because of N-terminal truncation, Hrad1B. Hrad1A
has 60% identity to the S. pombe rad1+
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 cancer cell lines. Chromosomal localization by
fluorescence in situ hybridization indicates that Hrad1 is located on chromosome 5p13.2-13.3. This region
is subject to loss of heterozygosity in several human cancers. Hrad1
also shares homology with the Saccharomyces cerevisiae
RAD17 and Ustilago maydis REC1 proteins. REC1 has
previously been characterized as a 3'
A Human Homologue of the Schizosaccharomyces pombe
rad1+ Checkpoint Gene Encodes an Exonuclease
,,,
Department of Experimental
Molecular Biology and the ¶ Department of Applied Molecular
Biology, Janssen Research Foundation, Turnhoutseweg 30, B-2340 Beerse, Belgium
5' exonuclease with a
C-terminal domain essential for cell cycle checkpoint function. We have
expressed and purified polyhistidine-tagged fusions of Hrad1A and
Hrad1B and show that HisHrad1A has 3' 5' exonuclease activity,
whereas HisHrad1B lacks such activity. The biological functions of the
two proteins remain to be determined.
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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