Proteins associated with the messenger ribonucleoprotein particle for the estrogen-regulated apolipoprotein II mRNA.
The stability of the mRNA for apolipoprotein (apo) II is regulated by estrogen [Gordon et al. (1988) J. Biol. Chem. 263, 2625-2631]. On the hypothesis tha estrogen regulation of apoII mRNA stability is mediated through mRNA-protein interaction, we have examined the messenger ribonucleoprotein particle (mRNP) for apoII mRNA following release from chicken liver polyribosomes. Polyribosomes containing undegraded apoII mRNA were obtained when tissue was homogenized without detergent, and polyribosomes were isolated following simultaneous addition of detergent and magnesium to a 20000g supernatant. ApoII mRNP released by EDTA sedimented at 12-18 S in sucrose gradients, and banded at rho = 1.4 g/mL in CsCl isopycnic centrifugation, indicative of a 3:1 ratio of protein to mRNA. A fraction in which apoII mRNP was enriched to 40-50% of total mRNP was prepared by successive size fractionation steps on sucrose gradients. Proteins associated with sucrose gradient enriched apoII mRNP were examined by iodination of UV-cross-linked proteins followed by SDS-polyacrylamide gel electrophoresis. Comparisons of proteins in highly enriched apoII mRNP to proteins in mRNP from non-estrogen-treated rooster liver did not reveal any differences. This result suggests that the major proteins associated with apoII mRNA are mRNP proteins also associated with the bulk of liver mRNAs.
MacDonald
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Department of Pharmacological Sciences, State University of New York, Stony Brook 11794-8651.
Williams
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D000818:Animals; D001053:Apolipoproteins / Q000235:genetics*; D002499:Centrifugation, Density Gradient; D002645:Chickens; D003432:Cross-Linking Reagents; D015342:DNA Probes; D004355:Drug Stability; D004492:Edetic Acid; D004967:Estrogens / Q000494:pharmacology; D008099:Liver / Q000737:chemistry; D008297:Male; D009693:Nucleic Acid Hybridization; D011132:Polyribosomes / Q000737:chemistry; D011498:Protein Precursors / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism*; D012261:Ribonucleoproteins / Q000378:metabolism*; D014466:Ultraviolet Rays
D001053:Apolipoproteins; D003432:Cross-Linking Reagents; D015342:DNA Probes; D004967:Estrogens; D011498:Protein Precursors; D012333:RNA, Messenger; D012261:Ribonucleoproteins; C054949:apolipoprotein II; C026452:messenger ribonucleoprotein; D004492:Edetic Acid