Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Schladebeck_Whi3_Ploidy

ABSTRACT: We analyzed and classified Whi3-regulated and ploidy-regulated genes in haploid and diploid strains of the Sigma1278b genetic background under vegetative growth conditions.

For this purpose, we measured transcriptional profiles of two different haploid MATa and one diploid MATa/a yeast strains of the following genotypes: WHI3 strain (SS_YHUM468=YHUM0468), whi3 strain (SS_ySS137=YHUM1920) and whi3-delta/whi3-delta strain (SS_ySS137dipl=YHUM2152). All three strains were grown in duplicate in YNB medium supplemented with tryptophan and uracil at 30 degrees C to an optical density of 1.0 before extraction of total RNA and transcriptional profiling

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Sarah Schladebeck

PROVIDER: E-MEXP-3610 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

The RNA-binding protein Whi3 is a key regulator of developmental signaling and ploidy in Saccharomyces cerevisiae.

Schladebeck Sarah S Mösch Hans-Ulrich HU

Genetics 20130614 1

In Saccharomyces cerevisiae, the RNA-binding protein Whi3 controls cell cycle progression, biofilm formation, and stress response by post-transcriptional regulation of the Cdc28-Cln3 cyclin-dependent protein kinase and the dual-specificity protein kinase Yak1. Previous work has indicated that Whi3 might govern these processes by additional, yet unknown mechanisms. In this study, we have identified additional effectors of Whi3 that include the G1 cyclins Cln1/Cln2 and two known regulators of biof ...[more]

PMID: 23770701

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Project description:The screen for genes involved in the spontaneous loss of heterozygosity mutagenesis (SLM) in diploid cells was done on the collections of deletion strains of Saccharomyces cerevisiae created by the Saccharomyces Genome Deletion Project (http://www-sequence.stanford.edu/group/yeast_deletion_project/). Screens were performed on the pools of clones. Barcode microarrays were used to detect the increased SLM frequency leading to relative increase in abundance of deletion clone in a pool. Collections used were: Homozygous Diploid (HD) and Essential (E) mixed to create single HDE pool. Three mutagenesis markers were used: (1) naturally existing mating type locus, and two newly introduced markers, (2) CAN1/can1delta, (3) URA3/ura3delta. 1. To detect the increased SLM frequency at the mating type locus HDE pool was crossed to MATa or MATalpha strain. Deletion clones that gained mating competence due to loss of heterozygosity (LOH) at the mating type locus gave rise to clones with the ability to grow on minimal medium. The relative abundance of these clones was compared to the abundance in the original HDE pool. 2. To detect the increased SLM frequency at the CAN1/can1delta locus the derivative HDE pool was created. Every cell of this pool had one of two copies of CAN1 gene deleted. The increased SLM frequency led to increased relative abundance of respective deletion clone under canavanine selection and could be estimated by comparing it to the abundance when pool was grown without selection. 3. To detect the increased SLM frequency at the URA3/ura3delta locus the derivative HDE pool was created. Every cell of this pool had one of two copies of URA3 gene deleted. The increased SLM frequency led to increased relative abundance of respective deletion clone under 5M-^R-fluoroorotic acid selection and could be estimated by comparing it to the abundance when pool was grown without selection. 4. For screens of SLM at CAN1 and URA3 markers the resistance control experiments were performed to detect genes whose deletion is sufficient to enable yeast cells with functional CAN1 or URA3 to grow in the presence of canavanine or 5M-^R- fluoroorotic acid respectively. 5. To include in the analysis slow-growing deletion clones control hybridizations were performed to detect genes whose deletion extends the doubling time of yeast cells.

Dataset Information

Schladebeck_Whi3_Ploidy

Publications

The RNA-binding protein Whi3 is a key regulator of developmental signaling and ploidy in Saccharomyces cerevisiae.

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