Copyright © 1989 Published by Elsevier Science B.V.
The C-terminal part of a gene partially homologous to CDC25 gene suppresses the cdc25-5 mutation in Saccharomyces cerevisiae
Received 15 September 1988;
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Abstract
In Saccharomyces cerevisiae, the product of the CDC25 gene is required for progression in the cell division cycle. It is necessary for cAMP production. It has been suggested that the CDC25 gene product acts through Ras proteins. We report the cloning of a DNA fragment from a new gene able to suppress the thermosensitive phenotype of the cdc25−5 mutation. It is unable to suppress the defect of a mutant of the adenylate cyclase gene or of the ras1, ras2ts double mutant. This DNA fragment prevents the drop in cAMP level in cdc25-5 mutant cells shifted to restrictive temperature. The complementing part of this fragment contains a truncated open reading frame (ORF) corresponding to the 3' end of a gene we named SCD25. The 584-amino acid sequence deduced from this ORF shares 45% identity with the 592-aa C-terminal part of the CDC25 ORF which is sufficient for complementation of cdc25 mutations. Some of the common sequences between these two genes are also partially homologous with the amino acid sequence of LTE1, another gene of S. cerevisiae. The capacity of the SCD25 fragment to suppress a cdc25 mutation and its homology to the C-terminal part of the CDC25 led us to propose that the CDC25 and the SCD25 C-terminal fragments each encode a protein domain which is capable in itself to support a similar biochemical function.
Author Keywords: Recombinant DNA; nucleotide sequence; cAMP; yeast; cell division; Ras activation; shuttle vector
Abbreviations: aa, amino acid(s); bp, base pair(s); cAMP, cyclic AMP; kb, kilobase(s) or 1000 bp; nt, nucleotide(s); ORF, open reading frame; SDS, sodium dodecyl sulfate; ss, single strand(ed); SSC, 0.15 M NaCl/0.015 M Na3 · citrate pH 7.6; ts, thermosensitive; wt, wild type; YEPD and YNB, see MATERIALS AND METHODS, section a







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