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Insights into the structure of the CCR4-NOT complex by electron microscopy.

FEBS letters | 2011

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.

Pubmed ID: 21669201 RIS Download

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Associated grants

  • Agency: Wellcome Trust, United Kingdom
    Id: 087658
  • Agency: NCRR NIH HHS, United States
    Id: P41 RR001081
  • Agency: NCRR NIH HHS, United States
    Id: P41 RR-0108
  • Agency: Wellcome Trust, United Kingdom
    Id: WT087658MA

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UCSF Chimera (tool)

RRID:SCR_004097

A highly extensible program for interactive visualization and analysis of molecular structures and related data, including density maps, supramolecular assemblies, sequence alignments, docking results, trajectories, and conformational ensembles. High-quality images and animations can be generated. Chimera includes complete documentation and several tutorials, and can be downloaded free of charge for academic, government, non-profit, and personal use.

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IMAGIC (tool)

RRID:SCR_014447

An image analysis software that can process spectra and other multi-dimensional data-sets. The software package is aimed at processing large data sets from (cryo-) electron microscopy, especially in the field of single particle analyses. This software can be used with light and raster-tunneling microscopes, computer tomographs, FT-IR spectrometers and other signal collecting devices. This resource provides three-dimensional data processing and angular reconstitution modules that allow the three-dimensional reconstruction with point-group symmetry from the two dimensional electron microscopy projections. These models aid in the analysis of the macromolecules.

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