The 5'-end structure of ovalbumin mRNA in isolated nuclei and polysomes.
We used the chemical reagents dimethylsulfate and 4'-aminomethyl-4,5',8-trimethylpsoralen and the enzyme T1 ribonuclease to compare the 5'-end structure of ovalbumin mRNA in situ in purified hen oviduct nuclei and polysomes with that of the isolated mRNA. The qualitative pattern of structure-dependent base modifications and T1 ribonuclease cleavage sites in intranuclear and polysomal ovalbumin mRNAs was found to be nearly identical to those in isolated ovalbumin mRNA. These structural data are consistent with the presence of a trigonal stem-loop structure at the 5'-end of ovalbumin mRNA (hairpin-1) in nuclei and polysomes. Similar results were obtained for a coding region structure (hairpin-3) in intranuclear ovalbumin mRNA. We have recently shown that hairpin-1 positively affects the rate of ovalbumin mRNA translation in vitro and is part of a high affinity binding site for eucaryotic initiation factor-2 (eIF-2). The presence of hairpin-1 in ovalbumin mRNA in both a pretranslation state (nuclei) and active translation state (polysomes) is consistent with its hypothesized biological function as an intracellular initiation signal that facilitates the translation of this mRNA.
Department of Chemistry, Hendrix College, Conway, AR 72032.
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Kopper
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Stallcup
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Hufford
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Liarakos
D000225:Adenine / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002467:Cell Nucleus / Q000737:chemistry*; D002645:Chickens; D003062:Codon; D003562:Cytidine / Q000378:metabolism; D005260:Female; D008745:Methylation; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010047:Ovalbumin / Q000235:genetics*; D010777:Photochemistry; D011132:Polyribosomes / Q000737:chemistry*; D012333:RNA, Messenger / Q000737:chemistry* / Q000378:metabolism; D006163:Ribonuclease T1 / Q000378:metabolism; D014307:Trioxsalen / Q000031:analogs & derivatives / Q000737:chemistry
D003062:Codon; D012333:RNA, Messenger; D003562:Cytidine; D010047:Ovalbumin; C018822:aminomethyltrioxsalen; D006163:Ribonuclease T1; D000225:Adenine; D014307:Trioxsalen