Gene, 105 (1991) 139-140 0 1991 Elsevier Science Publishers

GENE

B.V. All rights reserved.

139

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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D. Schild and R.K.M., unpublished results), suggesting that those two genes may act in part by stabilizing a repair complex of other RAD genes (Lovett and Mortimer, 1987).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,

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search

A: PROSITE:

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mutants

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S., Buratowski,

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(1989) 5718-5722. Henikoff,

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strength

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ing of yeast recombination repair genes and evidence that several are nonessential genes. In: Friedberg, E.C. and Bridges, B.A. (Eds.), Cellular Responses

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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