Insights into the structure of the CCR4-NOT complex by electron microscopy.
The CCR4-NOT complex is a deadenylation complex, which plays a major role for mRNA stability. The complex is conserved from yeast to human and consists of nine proteins NOT1-NOT5, CCR4, CAF1, CAF40 and CAF130. We have successfully isolated the complex using a Protein A tag on NOT1, followed by cross-linking ... on a glycerol gradient. All components of the complex were identified by mass spectrometry. Electron microscopy of negatively stained particles followed by image reconstruction revealed an L-shaped complex with two arms of similar length. The arms form an accessible cavity, which we think could provide an extensive interface for RNA-deadenylation.
Mesh Terms:
Cell Cycle Proteins, Humans, Mass Spectrometry, Microscopy, Electron, Models, Molecular, Multiprotein Complexes, Protein Subunits, RNA, Messenger, Ribonucleases, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Transcription Factors
Cell Cycle Proteins, Humans, Mass Spectrometry, Microscopy, Electron, Models, Molecular, Multiprotein Complexes, Protein Subunits, RNA, Messenger, Ribonucleases, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Transcription Factors
FEBS Lett.
Date: Jul. 21, 2011
PubMed ID: 21669201
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