CPEB Deficiency Stimulates PTEN and Stat3 mRNA Translation and Induces Hepatic Insulin Resistance
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
SUBMITTER: Mei Xu
PROVIDER: E-GEOD-28106 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:Analysis of CPEB translational regulator target mRNAs
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Project description:FOG-1/CPEB and FOG-3/Tob are the terminal regulators of the sex determination in C. elegans germ cells. CPEB and Tob proteins are both translational regulators. To investigate how FOG-1 and FOG-3 regulate germ cell sex determination we sought to identify the target mRNAs. We used transgenic epitope tagged animals (3xMyc::FOG-1 and FOG-3::3xFLAG). To identify the mRNA targets of FOG-1/CPEB and FOG-3/Tob on a genome wide scale we used RNA immunoprecipitation followed by microarray analysis. We found 81 putative mRNA targets of FOG-1 and 722 putative targets of FOG-3. 76 target mRNAs were common to both FOG-1 and FOG-3.
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Project description:FOG-1/CPEB and FOG-3/Tob are the terminal regulators of the sex determination in C. elegans germ cells. CPEB and Tob proteins are both translational regulators. To investigate how FOG-1 and FOG-3 regulate germ cell sex determination we sought to identify the target mRNAs. We used transgenic epitope tagged animals (3xMyc::FOG-1 and FOG-3::3xFLAG). To identify the mRNA targets of FOG-1/CPEB and FOG-3/Tob on a genome wide scale we used RNA immunoprecipitation followed by microarray analysis. We found 81 putative mRNA targets of FOG-1 and 722 putative targets of FOG-3. 76 target mRNAs were common to both FOG-1 and FOG-3. FOG-1 RNA-coimmunoprecipitation (RIP) samples were prepared as follows. Parallel RIPs were performed with wild type animals as the control IP. Worm extracts were made from wildtype and 3xMyc::FOG-1 late L3/early L4 animals. Myc affinity gel was used to immunoprecipitate 3xMyc::FOG-1 from the extracts. RNA was extracted from the pellets and analyzed on Affymetrix microarrays. 7 biological replicates of both 3xMyc::FOG-1 and wildtype were used. FOG-3 RNA-coimmunoprecipitation (RIP) samples were prepared as follows. Parallel RIPs were performed with wild type animals as the control IP. Worm extracts were made from wildtype and FOG-3::3xFLAG late L3/early L4 animals. Myc affinity gel was used to immunoprecipitate FOG-3::3xFLAG from the extracts. RNA was extracted from the pellets and analyzed on Affymetrix microarrays. 7 biological replicates of both FOG-3::3xFLAG and wildtype were used.
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Project description:The heat shock protein 90 (Hsp90) chaperone functions as a protein-folding buffer and plays a unique role promoting the evolution of new heritable traits. To better understand how Hsp90 can affect mRNA translation we screened more than 1600 factors involved in mRNA regulation for physical interactions with Hsp90 in human cells. The mRNA binding protein CPEB2 strongly binds Hsp90 via its prion domain. In a yeast model, transient inhibition of Hsp90 resulted in persistent activation of a CPEB translation reporter even in the absence of exogenous CPEB that persisted for 30 generations after the inhibitor was removed. Ribosomal profiling revealed that some endogenous yeast mRNAs, including HAC1, show a persistent change in translation efficiency following transient Hsp90 inhibition. Thus, transient loss of Hsp90 function can promote a non-genetic inheritance of a translational state affecting specific mRNAs, introducing a new mechanism by which Hsp90 can promote phenotypic variation.
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