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.
Copyright © 2011 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
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