Comprehensive analysis of diverse ribonucleoprotein complexes.
The 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.
Oeffinger
Marlene
M
Rockefeller University, 1230 York Avenue, New York, New York 10021, USA.
Wei
Karen E
KE
Rogers
Richard
R
DeGrasse
Jeffrey A
JA
Chait
Brian T
BT
Aitchison
John D
JD
Rout
Michael P
MP
D004268:DNA-Binding Proteins / Q000032:analysis / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D007163:Immunosorbent Techniques; D013058:Mass Spectrometry; D009687:Nuclear Proteins / Q000032:analysis / Q000235:genetics; D029543:Nucleocytoplasmic Transport Proteins / Q000032:analysis / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D018272:Porins; D039381:RNA Cap-Binding Proteins; D012331:RNA, Fungal / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis; D012335:RNA, Ribosomal / Q000032:analysis; D016601:RNA-Binding Proteins / Q000032:analysis / Q000235:genetics; D012261:Ribonucleoproteins / Q000032:analysis* / Q000235:genetics; D012269:Ribosomal Proteins / Q000032:analysis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis* / Q000235:genetics; D013205:Staphylococcal Protein A / Q000032:analysis / Q000235:genetics
D004268:DNA-Binding Proteins; C106898:MEX67 protein, S cerevisiae; D009687:Nuclear Proteins; D029543:Nucleocytoplasmic Transport Proteins; D018272:Porins; D039381:RNA Cap-Binding Proteins; D012331:RNA, Fungal; D012333:RNA, Messenger; D012335:RNA, Ribosomal; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; D012269:Ribosomal Proteins; C103382:SAC3 protein, S cerevisiae; C471175:SHE2 protein, S cerevisiae; C101398:STO1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D013205:Staphylococcal Protein A; C462947:Thp2 protein, S cerevisiae; C105887:YRA1 protein, S cerevisiae; C026452:messenger ribonucleoprotein; D049452:Green Fluorescent Proteins