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Genomic Run On (GRO): determination of the nascent transcriptional rates in a series of a mex67 mutant.

ABSTRACT: The goal of the project was to check the effect on transcription rate of a sudden depletion of Mex67 protein by using a thermosensitive mutant: mex67-5

ORGANISM(S): Saccharomyces cerevisiae S288C Saccharomyces cerevisiae

PROVIDER: GSE65228 | GEO | 2015/12/31

SECONDARY ACCESSION(S): PRJNA273450

REPOSITORIES: GEO

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Genomic Run On (GRO): determination of the nascent transcriptional rates in a series of a mex67 mutant.

Project description:The goal of the project was to check the effect on transcription rate of a sudden depletion of Mex67 protein by using a thermosensitive mutant: mex67-5 Cells were grown in YPD at 26ÂºC to 0.5 OD600. At that time cells were shifted to 37ÂºC to induce Mex67 protein inactivation and recovered after, 0, 5, 20, 40, 60 and 150 min.

2015-12-31 | E-GEOD-65228 | biostudies-arrayexpress

Genomic Run On (GRO): determination of the nascent transcriptional rates in a series of Hrp1 mutants.

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Genomic Run On (GRO): determination of the nascent transcriptional rates in a series of Hrp1 mutants.

Project description:The goal of the project was to check the differential effect on transcription rate of a sudden degron-induced depletion of Hpr1 protein and its comparison towards the effect of a constitutive absence of Hpr1 protein in a deletion strain. Cells were grown in YP-Raffinose + 0.1 mM CuSO4 at 26ÂºC to 0.5 OD600. At that cells were spun-down and changed to YP-Galactose without CuSO4 for 30 min at 26ÂºC. Then cells were shifted to 37ÂºC to induce degron-mediated proteolysis and recovered after 30 min. Samples include wt, hpr1delta and hpr1-degron after 0 or 30 min growth at 30ÂºC.

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Genomic Run On (GRO): determination of the nascent transcriptional rates in a spt16 mutant

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Genomic Run On (GRO): determination of the nascent transcriptional rates in a spt16 mutant

Project description:We analyzed the effect of SPT16 on the transcription rates, mRNA stabilities and mRNA levels by doing GRO experiments in a thermosensitive spt16 point mutant (G132D) mutant at 35 ÂºC comparing with permisive temperature (30 ÂºC) and a wt treated in the same way. Cells (wt or spt16 mutant) were grown in SC-ura at 30 ÂºC to 0.25 OD600. At that time Î± factor was added and let to stand for 160 min in the same conditions. Then each sample (wt or mutant) was separated in two aliquots. One of them remain at 30ÂºC and the other was incubated at 35ÂºC. Both aliquots in the presence of Î± factor for 1h more. Finally, samples were processed for GRO experiment as described in Garcia-Marinez et al (2004). Three replicates of the whole experiment were done.

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Genomic Run On (GRO): determination of the nascent transcriptional rates and mRNA levels in several yeast mutants.

Project description:In order to maintain the appropriate level of mRNA it is necessary coordinate simultaneously all the steps along the mRNA life cycle. It has been shown that several factors act in the regulation of gene expression as global coordinators. Thus, some kind of information is transferred from the nucleus to the cytoplasm, imprinted in the mRNA. In this way, it is conceivable the existence of mechanisms that ensure the balance between mRNA synthesis and degradation through the information flow from the cytoplasm to the nucleus and vice versa, as a crosstalk among both process to ensure the proper mRNA homeostasis in the cell. In this study, we have used null mutants related with transcription and mRNA degradation and we find that the lack of those factors impacts both over transcription and RNA stability at genome wide levels to compensate the impaired effect produce by the loss of each studied factor. This study focus on the transcriptional activity of RNA pol II in yeast genes. S. cerevisiae cells grown in YPD or YPGal to exponential phase were subjected to Genomic Run On. Data were normalized by the ArrayStat software. Home-made macroarrays containing 6035 ORF were used.

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Genomic Run On (GRO): determination of the nascent transcriptional rates and mRNA levels in several yeast mutants.

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Genomic Run On (GRO): determination of the nascent transcriptional rates and mRNA levels in several yeast mutants (SAC3, SUS1, SRC1)

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Genomic Run On (GRO): determination of the nascent transcriptional rates and mRNA levels in several yeast mutants (PUB1 and NGR1)

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