Comprehensive analysis of diverse ribonucleoprotein complexes

TitleComprehensive analysis of diverse ribonucleoprotein complexes
Publication TypeJournal Article
Year of Publication2007
AuthorsOeffinger M, Wei KE, Rogers R, DeGrasse JA, Chait BT, Aitchison JD, Rout MP
JournalNat Methods
Date PublishedNov
KeywordsDNA-Binding Proteins/analysis/genetics, Electrophoresis, Polyacrylamide Gel, Green Fluorescent Proteins/analysis/genetics, Immunosorbent Techniques, Mass Spectrometry, Nuclear Proteins/analysis/genetics, Nucleocytoplasmic Transport Proteins/analysis/genetics, Oligonucleotide Array Sequence Analysis, Ribonucleoproteins/*analysis/genetics, Ribosomal Proteins/analysis/genetics, RNA, Fungal/analysis, RNA, Messenger/analysis, RNA, Ribosomal/analysis, RNA-Binding Proteins/analysis/genetics, Saccharomyces cerevisiae Proteins/*analysis/genetics, Saccharomyces cerevisiae/genetics/*metabolism, Staphylococcal Protein A/analysis/genetics
AbstractThe study of the dynamic interactome of cellular ribonucleoprotein (RNP) particles has been hampered by severe methodological limitations. In particular, the affinity purification of intact RNP complexes from cell lysates suffers from RNA degradation, loss of interacting macromolecules and poor overall yields. Here we describe a rapid affinity-purification method for efficient isolation of the subcomplexes that dynamically organize different RNP biogenesis pathways in Saccharomyces cerevisiae. Our method overcomes many of the previous limitations to produce large RNP interactomes with almost no contamination.
Short TitleNature methods
Alternate JournalNature methods