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

FEBS Lett. 2011 Jul 21;585(14):2182-6. doi: 10.1016/j.febslet.2011.05.071. Epub 2011 Jun 12.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / isolation & purification
  • Cell Cycle Proteins / ultrastructure*
  • Humans
  • Mass Spectrometry / methods
  • Microscopy, Electron / methods
  • Models, Molecular
  • Multiprotein Complexes / chemistry*
  • Multiprotein Complexes / isolation & purification*
  • Protein Subunits / chemistry*
  • Protein Subunits / genetics
  • Protein Subunits / isolation & purification
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Ribonucleases / genetics
  • Ribonucleases / isolation & purification
  • Ribonucleases / ultrastructure*
  • Saccharomyces cerevisiae / chemistry
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / isolation & purification
  • Saccharomyces cerevisiae Proteins / ultrastructure*
  • Transcription Factors / genetics
  • Transcription Factors / isolation & purification
  • Transcription Factors / ultrastructure*

Substances

  • CDC39 protein, S cerevisiae
  • Cell Cycle Proteins
  • Multiprotein Complexes
  • Protein Subunits
  • RNA, Messenger
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • CCR4 protein, S cerevisiae
  • Ribonucleases
  • POP2 protein, S cerevisiae