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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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RNA-seq analysis of the budding yeast set1∆ strain and prp45(1-169) set1∆ double mutant strain

ABSTRACT: Prp45 is a budding yeast NineTeen-Complex associated factor, which plays a role in pre-mRNA splicing. Because its human ortholog, SNW1/SKIP, is known to functionally and physically interact with factors involved in transcription elongation and chromatin modifications, it was suggested to be one of the coupling factors which functionally connect splicing and transcription. To determine whether yeast Prp45 has this function as well we tested the genetic interactions between prp45(1-169) allele and deletions of histone modifiers and transcription elongation regulators. RNA-seq analysis was performed to address the changes in transcription and/or splicing in strains deleted in set1, a histone methyltransferase responsible for H3K4 methylation, and the corresponding prp45(1-169) set1 double mutant. Total RNA was isolated by combining phenol-chlorophorm extraction with MasterPure Yeast RNA Purification Kit (Epicentre). Ribodepletion, library preparation and sequencing were performed by BGI Genomics.

INSTRUMENT(S): Illumina HiSeq 2500

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Petr Folk

PROVIDER: E-MTAB-5335 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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RNA-seq analysis of the budding yeast prp45(1-169) strain (a scarless truncation)

Project description:Prp45 is a budding yeast NineTeen-Complex associated factor, which plays a role in pre-mRNA splicing. prp45(1-169) is a temperature-sensitive truncation allele expressing only the first 169 amino acids (the full-length protein consists of 379 amino acids). RNA-seq analysis was performed to determine changes in transcription and splicing in cells with prp45(1-169) allele prepared by the CRISPR/Cas9 method as a scarless truncation, i.e. with no tags and with preserved endogenous 3' UTR. Total RNA was isolated by combining phenol-chlorophorm extraction with MasterPure Yeast RNA Purification Kit (Epicentre). Ribodepletion, library preparation and sequencing were performed by BGI Genomics.

2024-08-30 | E-MTAB-5334 | biostudies-arrayexpress

RNA-seq analysis of the budding yeast rad6 strain and prp45(1-169) rad6 double mutant strain

Project description:Prp45 is a budding yeast NineTeen-Complex associated factor, which plays a role in pre-mRNA splicing. Because its human ortholog, SNW1/SKIP, is known to functionally and physically interact with factors involved in transcription elongation and chromatin modifications, it was suggested to be one of the coupling factors which functionally connect splicing and transcription. To determine whether the yeast Prp45 has this function as well we tested genetic interactions between prp45(1-169) allele and deletions of histone modifiers and transcription elongation regulators. RNA-seq analysis was performed to address the changes in transcription and/or splicing in strains deleted in rad6, a kinase phosphorylating serine 2 of the C-terminal domain of RNA Polymerase II and an E2 enzyme for H2BK123 monoubiquitination, respectively, and the corresponding double mutant with prp45(1-169). Total RNA was isolated by combining phenol-chlorophorm extraction with MasterPure Yeast RNA Purification Kit (Epicentre). Ribodepletion, library preparation and sequencing were performed by BGI Genomics.

2024-08-30 | E-MTAB-5336 | biostudies-arrayexpress

RNA-seq analysis of budding yeast wild type (BY4741) and prp45(1-169) mutant

Project description:Prp45 is a budding yeast NineTeen-Complex associated factor, which plays a role in pre-mRNA splicing. In addition to its documented involvement in the second step of splicing, here we show that it is important in the early stages of co-transcriptional spliceosome assembly as well. To determine the overall splicing efficiency of prp45 mutant cells and to reveal how the efficiency depends on the sequences which define introns (whether they are consensual or non-consensual), we performed RNA-seq analysis of prp45(1-169) and corresponding wild-type cells (two biological replicates each). Total RNA was isolated by combining phenol-chlorophorm extraction with MasterPure Yeast RNA Purification Kit (Epicentre). Ribodepletion, library preparation and sequencing were performed by BGI Genomics.

2017-02-10 | E-MTAB-5149 | biostudies-arrayexpress

Transcriptome analysis of budding yeast strains manipulated in RPL22A and RPL22B genes

Project description:Most Saccharomyces cerevisiae ribosomal protein genes (RPGs) are present as two paralogous copies that emerged during a whole genome duplication. The preservation of two copies of the same gene during the past ~100 million years enabled diversification of the regulation of their expression, and also partially of their functional properties. One example of such paralogous genes are RPL22A and RPL22B that are asymmetrically expressed and their protein sequences represent one of the most divergent pairs among budding yeast RPG paralogs. It was shown that introns in the RPL22A and RPL22B genes are important for regulation of their expression levels. To investigate functional differences between RPL22A and RPL22B we performed RNA-seq analysis of strains with complete deletions of either RPL22A or RPL22B, and also of strains with intron deletions in the RPL22 genes, in comparison with a wild-type strain. Cells were grown to mid-exponential phase in the rich YPAD medium. Total RNA was isolated by combining phenol-chlorophorm extraction with MasterPure Yeast RNA Purification Kit (Epicentre). Ribodepletion, library preparation and sequencing were performed by BGI Genomics.

2018-03-07 | E-MTAB-5275 | biostudies-arrayexpress

Cla4 catalyzed Set1 phosphorylation

Project description:H3K4me3 is catalyzed by the Set1/MLL family of methyltransferases, whose function in catalyzing H3K4me3 is unique. Impaired function of Set1/MLL family members can lead to many abnormalities, such as bone and nerve defects, leukemia, and even death. Although the Set1 family plays an important regulatory role in various biological processes, it is still unclear how the Set1 protein itself is regulated and how protein levels are maintained. Due to the numerous homologues, complex composition, and high molecular weight of Set1 in higher organisms, especially humans, related research is greatly limited. In brewing yeast, Set1 is the only methyltransferase that catalyzes H3K4me3 and is highly conserved between species. Therefore, yeast is an ideal model for studying the functions and mechanisms of the Set1 family. In addition, Set1 protein plays an important role in regulating gene transcription, promoting telomere silencing, and maintaining cell lifespan. The Set1 family also plays an important regulatory role in the occurrence and development of various cancers.

2023-10-24 | PXD042010 | Pride

Set1 and Set3

Project description:Set1 and Set3

| PRJNA638133 | ENA

Transcriptome analysis of yeast methyltransferase mutants set5∆, set1∆ and set5∆set1∆

Project description:Purpose: Understand the synergistic relationship between the methyltransferases Set1 and Set5 in the regulation of gene expression. Methods: Total mRNA was obtained from two independent biological replicates each of wildtype (WT), set1∆, set5∆, SET5 Y402A and set1∆set5∆ S. cerevisiae strains. Libraries were generated and sequenced using an Illumina HiSeq2000 platform. The sequence reads that passed quality filters were mapped using TopHat and expression levels were quantified using Cufflinks. Results: We generated FPKM expression values for each transcript and identified the differentially expressed genes using an FDR-adjusted p-value of 0.05. Subsequent data analysis was restricted to genes with fold-change greater than 1.7 relative to WT. Our results show that Set1 and Set5 have roles primarily in transcription repression. Moreover, lack of both Set1 and Set5 results in a synergistic exhacerbation of the transcriptional derepression observed in the single mutants. Further analysis revealed a specific enrichment of the Set5/Set1-repressed genes near repetitive DNA sequences of the genome. Conclusions: Our study uncovers an unexpected synergistic role of Set1 and Set5 in transcription repression of telomeric regions and Ty retrotransposons. mRNA profiles of wildtype (WT), set1∆, set5∆, SET5 Y402A and set1∆set5∆ were generated by sequencing using an Illumina HiSeq2000 platform. Two biological replicates of each strain were used.

2014-01-15 | E-GEOD-52086 | biostudies-arrayexpress

Heterochromatin assembly and transcriptome repression by Set1 in coordination with a class II histone deacetylase

Project description:Gene expression profiling of S. pombe set1/COMPASS/H3K4 mutants, atf1 and set1 atf1 deletion mutants, set1 clr3 deletion mutants; ChIP-chip tiling microarray profiling of S. pombe Set1, Atf1, H3K4me3 in wt/atf1 deletion mutants, H3K9me2 in wt/set1 clr3 deletion mutants Custom Agilent 60mer microarrays were used to assay gene expression in set1/COMPASS mutants in combination with atf1 and clr3 mutants, and to profile genome-wide binding of Set1, Atf1, H3K4me3 and H3K9me2 in S. pombe cells (two-color mutant vs. wildtype, ChIP vs. input experiments).

2014-12-12 | E-GEOD-63301 | biostudies-arrayexpress

Transcriptome analysis of yeast methyltransferase mutants set5∆, set1∆ and set5∆set1∆

2014-01-15 | GSE52086 | GEO

Transcription profiling of mouse ATCC CRL1606 hybridoma cells to evaluate microarray gene identification methods using spike-in experiments

Project description:Due to the lack of controlled microarray experimental data, evaluation of gene identification methods has been performed using simulated/manipulated data. As the ability of these approaches to mimic actual experimental data is questionable, conclusions drawn can be misleading. We applied spike-in approaches to develop experimental benchmark data and used it to evaluate various gene identification methods. mRNA was obtained from mouse ATCC CRL1606 hybridoma cells. From self hybridization results 200 genes that consistently exhibited a log ratio value close to 0 were randomly selected across a range of intensity. In-vitro transcripts of these 200 genes were obtained from their respective clones. From RNA gel electrophoresis and test array results, 169 of the 200 transcripts were deemed successful. Experiments were then conducted by spiking these 169 transcripts to the common pool of mRNA extracted from the hybridoma cells. With exception of the spiked in genes, the experimental set-up is identical to a self hybridization. This experiment is thus different from other microarray experiments as the set of differentially expressed genes is known as they have been artificially created.

2005-03-17 | E-MEXP-280 | biostudies-arrayexpress

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