Project description:The h+ and h- haploid cells of S. pombe mixtures flocculated and formed diploid cells when cultured in carbon limited YEPME medium. We first cultured S. pombe h+ YHL6381 and h- SPQ-01 cells respectively in YEPD medium and then mixed them at an 1:1 ratio and further cultured in YEPME medium. Sexual flocculation occured. We used microarrays to detail the the global programme of gene expression of flocculated YHL6381 and SPQ-01 mixtures cultured in YEPME, compared with the non flocculated cells mixtures cultured in YEPD, and identified distinct classes of up-regulated genes during this process.
Project description:The h+ and h- haploid cells of S. pombe mixtures flocculated and formed diploid cells when cultured in carbon limited YEPME medium. We first cultured S. pombe h+ YHL6381 and h- SPQ-01 cells respectively in YEPD medium and then mixed them at an 1:1 ratio and further cultured in YEPME medium. Sexual flocculation occured. We used microarrays to detail the the global programme of gene expression of flocculated YHL6381 and SPQ-01 mixtures cultured in YEPME, compared with the non flocculated cells mixtures cultured in YEPD, and identified distinct classes of up-regulated genes during this process. We first cultured S. pombe h+ YHL6381 and h- SPQ-01 cells respectively in YEPD medium and then mixed them at an 1:1 ratio and further cultured in a new medium for 90min. The flocculated cells mixtures cultured in YEPME named YSD, while the non flocculated cells mixtures cultured in YEPD named YSM.
Project description:The regulation of flocculation, surface adhesion and invasive growth in the fission yeast Schizosaccharomyces pombe has focused primarily at the transcriptional level, but little is known with regards to posttranscriptional control. Here, we identified the Pumilio protein Pfr1 as a novel posttranscriptional regulator of these processes. Deletion of pfr1+ prevented flocculation, surface adhesion and invasive growth under inducing conditions, while overexpression of pfr1+ was sufficient to trigger flocculation. The flocculent phenotype of pfr1+ overexpression was dependent on the presence of the Gsf2 flocculin, but not on the Mbx2, Cbf12 and Adn3 transcription factors. In addition, we used RNA immunoprecipitation and expression microarrays to identify pvg1+ and SPBPB7E8.01, which encode a galactose pyruvyltransferase and glycophosphatidylinositol membrane protein, respectively, as putative mRNA targets potentially degraded by Pfr1. The mRNAs of these genes were upregulated and downregulated in the pfr1 deletion and overexpression strains, respectively, and contained putative binding sites in the 3’-untranslated region. We also discovered that ccr4+ and ste13+, which encode components of the mRNA decay machinery, were required for these processes, but did not suppress the pfr1+ overexpression flocculent phenotype when deleted. This data suggest that these processes in S. pombe involve multiple posttranscriptional-regulatory pathways of which one requires Pfr1.
Project description:Transcriptional profiling and ChIP-chip analyses of S. pombe strains exhibiting flocculation phenotype by overexpression and deletion
Project description:Posttranscriptional Control of Flocculation, Surface Adhesion and Invasive Growth by the Pumilio gene pfr1+ in Schizosaccharomyces pombe
Project description:Puf3 is a RNA-binding protein, a member of the conserved Puf-protein family. Combining different functional genomics data, we have analyzed the role of Puf3 in post-transcriptional gene regulation in S. pombe. We present data on Puf3 interacting proteins and regulatory mRNA targets.
Project description:The regulation of flocculation, surface adhesion and invasive growth in the fission yeast Schizosaccharomyces pombe has focused primarily at the transcriptional level, but little is known with regards to posttranscriptional control. Here, we identified the Pumilio protein Pfr1 as a novel posttranscriptional regulator of these processes. Deletion of pfr1+ prevented flocculation, surface adhesion and invasive growth under inducing conditions, while overexpression of pfr1+ was sufficient to trigger flocculation. The flocculent phenotype of pfr1+ overexpression was dependent on the presence of the Gsf2 flocculin, but not on the Mbx2, Cbf12 and Adn3 transcription factors. In addition, we used RNA immunoprecipitation and expression microarrays to identify pvg1+ and SPBPB7E8.01, which encode a galactose pyruvyltransferase and glycophosphatidylinositol membrane protein, respectively, as putative mRNA targets potentially degraded by Pfr1. The mRNAs of these genes were upregulated and downregulated in the pfr1 deletion and overexpression strains, respectively, and contained putative binding sites in the 3â-untranslated region. We also discovered that ccr4+ and ste13+, which encode components of the mRNA decay machinery, were required for these processes, but did not suppress the pfr1+ overexpression flocculent phenotype when deleted. This data suggest that these processes in S. pombe involve multiple posttranscriptional-regulatory pathways of which one requires Pfr1. We generated 2 overexpression microarrays with dye swap that were biological replicates, 2 deletion microarrys with dye swap. Mutants samples were compared to empty vector control or wild type. 1 RIP-chip array was generated with IP rna compared to total RNA from the sample.