Isolation of messenger-like ribonucleoproteins.
Subribosomal and polyribosomal messenger ribonucleoproteins (mRNPs) were isolated from Ehrlich ascites tumor cells by a method involving sedimentation of polyribosomal and subribosomal particles, dissociation with EDTA, and rate-zonal sedimentation. The fractions containing mRNA protein particles were applied to glass fiber filters and extensively washed with buffer containing 0.5 M KCl. The eluted material was demonstrated to be an RNA-protein complex containing poly(A)-rich RNA, heterogeneous in size, and free of 18S or 28S rRNA. mRNA function for the RNA was suggested by its ability to direct protein synthesis in a cell-free protein-synthesizing system derived from wheat germ embryos. Analysis of the proteins associated with subribosomal and polyribosomal mRNPs by iodination and sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed at least seven similar proteins. The apparent molecular weights of the three most prominent proteins were 78,000, 52,000, and 34,000. Analysis of reticulocyte polyribosomal mRNPs revealed an increased prominence of the 78,000 and 52,000 molecular weight proteins relative to the other protein bands.
D000818:Animals; D002286:Carcinoma, Ehrlich Tumor / Q000032:analysis* / Q000378:metabolism; D002458:Cell Fractionation; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D051379:Mice; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000096:biosynthesis; D009698:Nucleoproteins / Q000302:isolation & purification* / Q000378:metabolism; D011132:Polyribosomes / Q000032:analysis; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000302:isolation & purification* / Q000378:metabolism; D012334:RNA, Neoplasm / Q000302:isolation & purification* / Q000378:metabolism; D012335:RNA, Ribosomal / Q000302:isolation & purification* / Q000378:metabolism; D012270:Ribosomes / Q000032:analysis / Q000378:metabolism
D009363:Neoplasm Proteins; D009698:Nucleoproteins; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D012335:RNA, Ribosomal