Molecular Cloning and Transcriptional Analysis of the Start Gene CDC25 of Saccharomyces Cerevisiae

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1986 Sep 1

PMID 16453707

Citations 12

Authors

Enzo Martegani

Maurizio D Baroni

Gianni Frascotti

Lilia Alberghina

Affiliations

Soon will be listed here.

Abstract

To isolate the CDC25 gene of Saccharomyces cerevisiae we have transformed a cdc25-1, trp1 strain with a yeast gene bank constructed in YRp7 vector, selecting trp clones able to grow at restrictive temperature. From several independent positive clones we have recovered a plasmid, called pDGEm-1, that bears a 5-kb genomic fragment and is able to give a full complementation of the cdc25-1 mutation. The genomic sequence has been subcloned and a good complementation obtained with a 2-kb fragment. Several stable integrative trp, cdc transformants have been constructed. Their genetic and molecular analysis indicates that we have cloned the true CDC25 gene. Northern blot hybridization has revealed the presence of a 5-kb mRNA transcribed by the CDC25 gene. This mRNA is also present in nitrogen-starved cells and during the re-enter in cell cycle from starvation, suggesting a constitutive transcription. Transformants bearing the cloned sequence on multicopy plasmid and integrative transformants that bear the CDC25 gene, flanked by plasmid sequences, show an altered control of the cell cycle and fail to arrest in G1 unbudded phase in stationary phase conditions.

Citing Articles

Glucose signaling-mediated coordination of cell growth and cell cycle in Saccharomyces cerevisiae.

Busti S, Coccetti P, Alberghina L, Vanoni M Sensors (Basel). 2012; 10(6):6195-240.

PMID: 22219709 PMC: 3247754. DOI: 10.3390/s100606195.

Involvement of distinct G-proteins, Gpa2 and Ras, in glucose- and intracellular acidification-induced cAMP signalling in the yeast Saccharomyces cerevisiae.

Colombo S, Ma P, Cauwenberg L, Winderickx J, Crauwels M, Teunissen A EMBO J. 1998; 17(12):3326-41.

PMID: 9628870 PMC: 1170671. DOI: 10.1093/emboj/17.12.3326.

The C-terminal part of the CDC25 gene product has Ras-nucleotide exchange activity when present in a chimeric SDC25-CDC25 protein.

Boy-Marcotte E, Buu A, Soustelle C, Poullet P, Parmeggiani A, Jacquet M Curr Genet. 1993; 23(5-6):397-401.

PMID: 8319295 DOI: 10.1007/BF00312625.

Isolation and characterization of Schizosaccharomyces pombe mutants phenotypically similar to ras1-.

Fukui Y, Yamamoto M Mol Gen Genet. 1988; 215(1):26-31.

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Domains of the Saccharomyces cerevisiae CDC25 gene controlling mitosis and meiosis.

Munder T, Mink M, Kuntzel H Mol Gen Genet. 1988; 214(2):271-7.

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