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Global Identification of Noncoding RNAs in S. cerevisiae


ABSTRACT: Genome-wide detection of novel non-coding RNAs in S. cerevisiae by modulating an RNase P pathway through the depletion of a component RPP1. Nearly 400,000 36-mer oligonucleotide probes, tiling the entire yeast genome including the mitochondrial chromosome with an average gap of 10 bases between two consecutive probes, were synthesized on glass slides using a mask-less array synthesizer. RNA samples for hybridizing to the arrays were extracted from a conditional lethal allele of S. cerevisiae created by placing the RPP1 gene under control of GAL10 promoter. It allowed the expression of RPP1 in galactose-containing culture medium but suppressed its expression in glucose-containing medium. A wild-type isogenic strain was used as a control. Both strains were initially grown in galactose-containing medium and subsequently transferred and resuspended into glucose-containing medium. Eight arrays were hybridized with RNA extracted from the Rpp1-depleted cells at 0, 4, 7, 12, 16, 21, 30h, and the control cell at 30h, after initial transfer to glucose-containing medium.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Waraporn Tongprasit 

PROVIDER: E-GEOD-4275 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Global identification of noncoding RNAs in Saccharomyces cerevisiae by modulating an essential RNA processing pathway.

Samanta Manoj Pratim MP   Tongprasit Waraporn W   Sethi Himanshu H   Chin Chen-Shan CS   Stolc Viktor V  

Proceedings of the National Academy of Sciences of the United States of America 20060306 11


Noncoding RNAs (ncRNAs) perform essential cellular tasks and play key regulatory roles in all organisms. Although several new ncRNAs in yeast were recently discovered by individual studies, to our knowledge no comprehensive empirical search has been conducted. We demonstrate a powerful and versatile method for global identification of previously undescribed ncRNAs by modulating an essential RNA processing pathway through the depletion of a key ribonucleoprotein enzyme component, and monitoring d  ...[more]

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