Mechanisms of cell cycle control revealed by a systematic and quantitative overexpression screen in S. cerevisiae

• Published in: PLoS genetics 2008-07, Vol.4 (7), p.e1000120-e1000120
• Main Authors: Niu, Wei, Li, Zhihua, Zhan, Wenjing, Iyer, Vishwanath R, Marcotte, Edward M
• Format: Article
• Language: English
• Subjects: Basic-Leucine Zipper Transcription Factors - genetics; Basic-Leucine Zipper Transcription Factors - metabolism; Cell Biology/Gene Expression; Cell cycle; Cell Cycle - drug effects; Cell Cycle Proteins - genetics; Cell Cycle Proteins - metabolism; Cell division; Chromosome Segregation; DNA Damage; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Flow Cytometry; Gene Expression Regulation, Fungal - drug effects; Genes; Genetics and Genomics/Functional Genomics; Genetics and Genomics/Gene Expression; Genetics and Genomics/Gene Function; Genetics and Genomics/Genomics; Hydroxyurea - pharmacology; Molecular Biology/DNA; Mutation; Nocodazole - pharmacology; Nucleic Acid Synthesis Inhibitors - pharmacology; Oligonucleotide Array Sequence Analysis; Phenotype; Pheromones - metabolism; Proteins; Replication Protein A; Repressor Proteins - genetics; Repressor Proteins - metabolism; Saccharomyces cerevisiae - cytology; Saccharomyces cerevisiae - drug effects; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - physiology; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; Signal Transduction; Transcription Factors - genetics; Transcription Factors - metabolism; Tubulin Modulators - pharmacology; Yeast
• ISSN: 1553-7404; 1553-7390; 1553-7404
• DOI: 10.1371/journal.pgen.1000120

Regulation of cell cycle progression is fundamental to cell health and reproduction, and failures in this process are associated with many human diseases. Much of our knowledge of cell cycle regulators derives from loss-of-function studies. To reveal new cell cycle regulatory genes that are difficult to identify in loss-of-function studies, we performed a near-genome-wide flow cytometry assay of yeast gene overexpression-induced cell cycle delay phenotypes. We identified 108 genes whose overexpression significantly delayed the progression of the yeast cell cycle at a specific stage. Many of the genes are newly implicated in cell cycle progression, for example SKO1, RFA1, and YPR015C. The overexpression of RFA1 or YPR015C delayed the cell cycle at G2/M phases by disrupting spindle attachment to chromosomes and activating the DNA damage checkpoint, respectively. In contrast, overexpression of the transcription factor SKO1 arrests cells at G1 phase by activating the pheromone response pathway, revealing new cross-talk between osmotic sensing and mating. More generally, 92%-94% of the genes exhibit distinct phenotypes when overexpressed as compared to their corresponding deletion mutants, supporting the notion that many genes may gain functions upon overexpression. This work thus implicates new genes in cell cycle progression, complements previous screens, and lays the foundation for future experiments to define more precisely roles for these genes in cell cycle progression.