Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

RNA-affinity isolations 13 RBPs

ABSTRACT: To identify RNAs specifically associated with potential RBPs, yeast cells expressing TAP-tagged RBP or the wild-type strain BY4741 (mock control) were grown to mid-log phase in rich medium and harvested by centrifugation. RNA affinity isolations were essentially performed as described (Gerber et al. 2004 PLoS Biol.; see protocol). In brief, TAP-tagged protein were captured from cell extracts with IgG coupled agarose beads (Sigma) and released by incubation with a site-specific protease (AcTEV, Sigma). from the extract (input) and from the affinity isolates was purified with the RNeasy Mini/ Micro Kit (Qiagen). RNAs were reverse transcribed using a mixture of oligo-dT and random nonamer oligos in the presence of amino-allyl dUTP/dNTP mixture. cDNAs were fluorescently labeled and hybridized on yeast oligo microarrays over night at 42 degrees in formamide-based hybridization buffer. Antigenic peptide used in IP: TAP-tag An all pairs experiment design type is where all labeled extracts are compared to every other labeled extract.

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Andre Gerber

PROVIDER: E-GEOD-21864 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

A screen for RNA-binding proteins in yeast indicates dual functions for many enzymes.

Scherrer Tanja T Mittal Nitish N Janga Sarath Chandra SC Gerber André P AP

PloS one 20101111 11

Hundreds of RNA-binding proteins (RBPs) control diverse aspects of post-transcriptional gene regulation. To identify novel and unconventional RBPs, we probed high-density protein microarrays with fluorescently labeled RNA and selected 200 proteins that reproducibly interacted with different types of RNA from budding yeast Saccharomyces cerevisiae. Surprisingly, more than half of these proteins represent previously known enzymes, many of them acting in metabolism, providing opportunities to direc ...[more]

PMID: 21124907

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Dataset Information

RNA-affinity isolations 13 RBPs

Publications

A screen for RNA-binding proteins in yeast indicates dual functions for many enzymes.

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