Identification and analysis of a DNA fragment from Saccharomyces kluyveri that can complement the loss of CDC25 function in Saccharomyces cerevisiae.
In the budding yeast, Saccharomyces cerevisiae, the function of wild-type Ras proteins is dependent on the CDC25 protein, which promotes the exchange of guanine nucleotides bound to Ras. To facilitate the identification of proteins which similarly regulate Ras function in higher eukaryotes, we have identified the CDC25 gene from another budding yeast, Saccharomyces kluyveri, by low-stringency hybridization to an S. cerevisiae CDC25 restriction fragment. This protein, SKCDC25, shares significant amino acid homology with CDC25, SCD25, and Ste6 of Schizosaccharomyces pombe in the C-terminal portion of the protein. The expression of SKCDC25 in a temperature-sensitive cdc25 strain of S. cerevisiae complements the loss of endogenous CDC25 activity. The identification of the highly conserved C-terminal sequences, which direct bona fide CDC25 activity within these proteins, will aid in the isolation of CDC25 genes from higher eukaryotes.
Prigozy
T
T
Kenneth Norris Jr. Cancer Research Hospital, Department of Biochemistry, University of Southern California School of Medicine, Los Angeles 90033.
Gonzales
E
E
Broek
D
D
D018528:ATP-Binding Cassette Transporters; D000595:Amino Acid Sequence; D001483:Base Sequence; D018797:Cell Cycle Proteins; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000032:analysis / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D006023:Glycoproteins; D020662:Guanine Nucleotide Exchange Factors; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D012440:Saccharomyces / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D029702:Schizosaccharomyces pombe Proteins; D016415:Sequence Alignment; D020666:ras-GRF1
D018528:ATP-Binding Cassette Transporters; D018797:Cell Cycle Proteins; D004271:DNA, Fungal; D005656:Fungal Proteins; D006023:Glycoproteins; D020662:Guanine Nucleotide Exchange Factors; C489277:STE6 protein, S cerevisiae; C489278:STE6 protein, S pombe; D029701:Saccharomyces cerevisiae Proteins; D029702:Schizosaccharomyces pombe Proteins; D020666:ras-GRF1