Gene, 105 (1991) 139-140 0 1991 Elsevier Science Publishers
GENE
B.V. All rights reserved.
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06020
Nucleotide sequence of the RAD57 gene of Saccharomyces (DNA
repair;
meiosis;
recombinant
DNA;
recombinational
repair;
cevevisiae *
x-ray sensitivity;
yeast)
Jonathan A. Kans a* * and Robert K. Mortimer a*~ tl Department ofMolecular
and Cell Biology, University of California, Berkeley,
CA 94720 (U.S.A.),
and” Division of Cell
and Molecular Biolog),,
Lawrence Berkeley Laboratory, Berkeley, CA 94720 (U.S.A.) Received by J. Marmur: 23 May 1991 Accepted: 26 May 1991 Received at publishers: 27 June 1991
SUMMARY
We have determined the nucleotide (nt) sequence of the RAD57 gene of Saccharomyces reading frame of 1380 bp. The deduced amino acid sequence of 460 residues contains shows significant similarity to the preliminary sequence of RAD51.
The product of the RAD.57 gene of the yeast S. cerevisiae participates in the repair of x-ray-induced damage to DNA and in meiosis (Game and Mortimer, 1974; Game, 1983). The gene is in the RAD50-RAD57 epistasis group, and is thought to be involved in recombinational repair, since mutants of RAD57 eliminate the increased x-ray resistance of haploid cells when sister chromatids are present during DNA replication and in early G2 (Game and Mortimer, 1974). RAD57 mutants, like those of RAD55, are coldsensitive for x-ray damage repair (including null mutations; Correspondence to: Dr. R.K. Mortimer, Department of Molecular and Cell Biology, 102 Donner Laboratory, University of California, Berkeley, CA 94720 (U.S.A.) Tel. (415)642-8877; * On request, the sequencing ** Present
Fax (415)642-8589.
the authors results
address:
will supply detailed
experimental
evidence
for
in this brief note. National
National
Library
Rockville
Pike, Bethesda,
Center
of Medicine,
for Biotechnology
National
Institutes
Information,
of Health,
8600
kb, kilobase
or
MD 20894 (U.S.A.)
Tel. (301)496-2475. Abbreviations:
aa, amino
1000 bp; nt, nucleotide(s); RAD, gene(s) encoding
acid(s);
bp, base pair(s);
RAD, protein(s) RAD.
participating
in DNA repair;
cerevisiae.
a potential
contains an open nt-binding sequence and
RAD57
D. Schild and R.K.M., unpublished results), suggesting that those two genes may act in part by stabilizing a repair complex of other RAD genes (Lovett and Mortimer, 1987). The RAD57 gene has been cloned (Schild et al., 1983), and the cloned gene integrates at the known location of RAD57 on chromosome IV (D. Schild and R.K.M., unpublished results). We have subcloned the gene and sequenced its coding region. RAD57 encodes an open reading frame of 460 aa with a deduced M, of 52 898. Using the BLAST local alignment search tool (Altschul et al., 1990) we have found significant aa sequence similarity between RAD57 and the preliminary sequence of RADSl (M. Aker, G. Basile and R.K.M., unpublished results). Significant sequence similarity was not detected between RAD57 and any other yeast protein (M. Goebl, personal communication). Motif analysis using the PROSITE database (Bairoch, 1990) revealed that aa 125-132 match the nt-binding consensus aa sequence GXXXXGK[ S/T] (Walker et al., 1982) where X stands for any aa; this motif is also conserved in the preliminary sequence of RADSl. In addition, aa 221-225 match most of the nt-binding consensus aa sequence hhhhD[ D/E], where h stands for a hydrophobic aa (Higgins et al., 1985).
60
ACKNOWLEDGEMENTS
120
240
This work was supported by a grant from the Office of Health and Environmental Research of the U.S. Department of Energy under contract DE-AC03-76SF00098 and by U.S. Public Health Service grants GM30990, 5 P40 RR04231-02 and ES07075. We wish to thank Dr. David Schild for the initial restriction mapping and subcloning of RAD57, and for helpful comments on the manuscript. We also thank Dr. Mark Goebl for performing FASTA sequence analysis on unpublished yeast sequences. BLAST computation was performed at the NCBI using BLAST network service. REFERENCES .4ltschul,
S.F.,Gish,
W., Miller, W., Myers, E.W. and Lipman,
local alignment Bairoch,
search
A: PROSITE:
University
A Dictionary
of Geneva,
Spencer.
Ful3dament~
of Protein
Sixth Release,
Game, J.C.: Radiation-sensitive J.F.T..
mutants
Applied
Sites and Patterns.
1990. and repair in yeast. In: Spencer,
D.M. and Smith, and
D.J.: Basic
tool J. Mol. Bioi. 215 (1990) 403-410.
A.R.W.
Aspects.
(Eds.),
Yeast Genetics:
Springer-Verlag,
New
York,
1983, pp~ 109-137. Game, J.C. and Mortimer, R.K.: A genetic study of X-ray mutants in yeast. Mutation Res. 24 (1974) 281-292. Hahn,
S., Buratowski,
S., Sharp,
binding protein TFIID and nonconsensus
P.A. and Guarente,
binds to TATA elements
DNA sequences.
sensitive
L.: Yeast TATAwith both consensus
Proc. Natl. Acad.
Sci. USA 86
(1989) 5718-5722. Henikoff,
S., Kelly, J.D. and Cohen,
yeast distal to a control Higgins,
CF.,
binding
Hiles, I.D., Whalley,
by membrane
protein-dependent
E.H.: Transcription
sequence.
K. and Jamieson,
components
transport
Lovett, S.T. and Mortimer,
of bacterial
osmotic
strength
F., Nicklen,
systems.
ternlinating
and mating
inhibitors.
type. Genetics
5463-5467. Schild, D., Calderon,
Proc.
ofnull mutants
I.L., Contopoulou,
Acad.
of the
of temperature,
116 (1987) 547-553.
A.R.: DNA sequencing Natl.
binding
EMBO J. 4 (1985) 1033-1039.
R.K.: Characterization
S. and Coulson,
in
D.J.: Nucleotide
periplasmic
RAD.55 gene of Succharomyces cerevisiae: effects Sanger,
terminates
Cell 33 (1983) 607-614.
Sci.
with chain-
USA
C.R. and Mortimer,
74 (1977) R.K.: Clon-
ing of yeast recombination repair genes and evidence that several are nonessential genes. In: Friedberg, E.C. and Bridges, B.A. (Eds.), Cellular Responses
to DNA Damage.
Alan R. Liss, New York, 1983,
pp. 4 17-427. Walker,
J.E., Saraste,
related sequences
Fig. 1. Nucleotide
sequence
of the RADS7 gene and the deduced
aa
sequence. Sequencing was done by the dideoxy method of Sanger et al. (1977) using the Sequenas?’
chain-termination kit (United States
Biochemicals,
determined
Cleveland,
OH),
and
sequence
was
inde-
pendently for both strands. The nt 1789-1891 in the 3’-untranslated region were determined for only the coding strand. Two kinds of consensus nt sequences for transcription initiation (Hahn et al., 1989) and termination (Henikotfet al., 1983) are underlined. Stop codon is marked by a dot. Consensus aa sequences for nt binding (Walker et al., 1982; Higgins et al., 1985) are overscored. The nt sequence was submitted to the GenBank,
EMBL and DDBJ Nucleotide
the accession
No. M6506
1.
Sequence
Databases
under
M., Runswick,
M.J. and Gay,
in the G[-and /?-subunits
kinases
and other ATP-requiring
binding
fold. EMBO
enzymes
J. 1 (1982) 945-951.
N.J.: Distantly
of .4TP-synthase, and a common
myosin, nucleotide