Association of messenger ribonucleic acid with mammalian microsomal membranes: characterization by analysis of cell-free translation products
- 12 June 1979
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 18 (12) , 2622-2627
- https://doi.org/10.1021/bi00579a029
Abstract
Total rough microsomes isolated from the dog pancreas were stripped of membrane-bound polysomes by treatment with either EDTA or puromycin and 0.5 M KCl. The stripped microsomal membranes were isolated relatively free from contamination by buoyant density centrifugation, and mRNA was isolated from both the membrane fraction and the released material. Depending on the method used to strip the rough microsomes, a variable but small percentage (3-15%) of the cellular poly(A)-containing mRNA was attached to the microsomal membranes. Reextraction of isolated microsomal membranes with puromycin and 0.5 M KCl reduced the content of membrane-associated mRNA by approximately 50%, resulting in less than 2% of the total membrane-bound polysomal mRNA remaining associated with the microsomal membranes. The membrane-associated mRNA was characterized by translation in the wheat germ cell-free protein synthesizing system; the products were analyzed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The translation products of the membrane-associated mRNA were identical with those from the total pancreas mRNA and those obtained using mRNA isolated from material released directly from the rough microsomes.This publication has 14 references indexed in Scilit:
- Binding of viral glycoprotein mRNA to endoplasmic reticulum membranes is disrupted by puromycin.The Journal of cell biology, 1977
- Studies on the mechanism for entry of vesicular stomatitis virus glycoprotein g mRNA into membrane-bound polyribosome complexesThe Journal of cell biology, 1977
- Cell-free synthesis of fish preproinsulin, and processing by heterologous mammalian microsomal membranes.Proceedings of the National Academy of Sciences, 1977
- Functional interaction of plant ribosomes with animal microsomal membranesBiochemical and Biophysical Research Communications, 1977
- Retention of mRNA on the endoplasmic reticulum membranes after in vivo disassembly of polysomes by an inhibitor of initiation.The Journal of cell biology, 1976
- Direct association of messenger RNA labeled in the presence of fluoroorotate with membranes of the endoplasmic reticulum in rat liver.The Journal of cell biology, 1976
- Homology in amino-terminal sequence of precursors to pancreatic secretory proteins.Proceedings of the National Academy of Sciences, 1975
- [10] Assay of inorganic phosphate, total phosphate and phosphatasesPublished by Elsevier ,1966
- A SIMPLE METHOD FOR THE ISOLATION AND PURIFICATION OF TOTAL LIPIDES FROM ANIMAL TISSUESJournal of Biological Chemistry, 1957
- PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENTJournal of Biological Chemistry, 1951