Transcriptomics

Dataset Information

121

Chromatin immunoprecipitation with yeast She proteins


ABSTRACT: RNA co-immunoprecipitations with C-terminal protein A-tagged She proteins. One liter of cells were cultured at 30 degrees in YPAD medium and collected during exponential growth by centrifugation. Cells were washed twice and broken mechanically with glass beads. Extracts were incubated with IgG-agarose beads (Sigma). The beads were washed four times, and She proteins were released from the beads by cleavage with TEV-protease (Invitrogen). RNA was isolated by phenol/chloroform extraction and isopropanol precipitation from TEV eluates, which corresponds to the purified fraction, and from extracts (input). Both RNA samples, input and purified, were reverse transcribed and labeled with the fluorescent dyes Cy3 and Cy5 (Amersham), respectively. The samples were mixed and competitively hybridized to yeast DNA microarrays containing all yeast genes

ORGANISM(S): Saccharomyces cerevisiae  

SUBMITTER: Janos Demeter   Andre Gerber  

PROVIDER: E-SMDB-2092 | ArrayExpress | 2005-10-14

REPOSITORIES: ArrayExpress

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Publications

Widespread cytoplasmic mRNA transport in yeast: identification of 22 bud-localized transcripts using DNA microarray analysis.

Shepard K A KA   Gerber A P AP   Jambhekar A A   Takizawa P A PA   Brown P O PO   Herschlag D D   DeRisi J L JL   Vale R D RD  

Proceedings of the National Academy of Sciences of the United States of America 20030917 20


Cytoplasmic mRNA localization provides a means of generating cell asymmetry and segregating protein activity. Previous studies have identified two mRNAs that localize to the bud tips of the yeast Saccharomyces cerevisiae. To identify additional localized mRNAs, we immunoprecipitated the RNA transport components She2p, She3p, and Myo4p and performed DNA microarray analysis of their associated RNAs. A secondary screen, using a GFP-tagged RNA reporter assay, identified 22 mRNAs that are localized t  ...[more]

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