Unknown,Transcriptomics,Genomics,Proteomics

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Cryptic Transcription in a set2∆ in Saccharomyces cerevisiae


ABSTRACT: In cells lacking the histone methyltransferase Set2, initiation of RNA polymerase II transcription occurs inappropriately within the protein-coding regions of genes, rather than being restricted to the proximal promoter. Here, we mapped the transcripts produced in an S. cerevisiae strain lacking Set2, and applied rigorous statistical methods to identify sites of cryptic transcription at high resolution. Wild type (BY4741) and set2∆ (BY4741) strains were grown at 30°C in YPD (1% yeast extract, 2% peptone, 2% dextrose) to an OD600 of 0.6-0.8. For each of the three replicates, Total RNA was extracted by acid-phenol method (Xiao et al. 2003). Double-stranded cDNA was prepared using an Invitrogen SuperScript™ (Cat No. 11917-010) primed with Oligo(dt) and random hexamers. For each replicate, the wt and set2∆ cDNA were independetly fluorescently labeled and comparatively hybridized to high-resolution 385K Saccharomyces cerevisiae CGH arrays (2005-08-16_SCER_WG_CGH) with Tm-normalized probes. In one of the replicates, assignment of the fluorescent label was reversed.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Jason Lieb 

PROVIDER: E-GEOD-13310 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

The Set2/Rpd3S pathway suppresses cryptic transcription without regard to gene length or transcription frequency.

Lickwar Colin R CR   Rao Bhargavi B   Shabalin Andrey A AA   Nobel Andrew B AB   Strahl Brian D BD   Lieb Jason D JD  

PloS one 20090319 3


In cells lacking the histone methyltransferase Set2, initiation of RNA polymerase II transcription occurs inappropriately within the protein-coding regions of genes, rather than being restricted to the proximal promoter. It was previously reported that this "cryptic" transcription occurs preferentially in long genes, and in genes that are infrequently transcribed. Here, we mapped the transcripts produced in an S. cerevisiae strain lacking Set2, and applied rigorous statistical methods to identif  ...[more]

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