Abstract
Poliovirus RNA that has been derivatized at the 3'-end with NaIO(4)-NaB(3)H(4) yields, after hydrolysis with alkali or RNase T2, predominantly labeled residues of modified adenosine; no labeled nucleoside derivative is produced by digestion with RNase A or RNase T1. The 3'-terminal bases of the RNA are, therefore,...ApA(OH). Hydrolyzates of poliovirus [(32)P]RNA, after exhaustive digestion with RNase T1 or RNase A, contain, besides internal oligonucleotides, polynucleotides resistant to further action of ribonucleases T1 and A, respectively; these polynucleotides were isolated by membrane-filter binding or ion-exchange chromatography. The sequence of the T1-resistant polynucleotide was determined to be (Ap)(n)A(OH), that of the RNase A-resistant polynucleotide was GpGp(Ap)(n)A(OH). The chain length (n) of the polyadenylic acid, as analyzed by different methods, averages 89 nucleotides. Gel electrophoresis revealed heterogeneity of the size of poly(A). Poliovirus RNA, when labeled in vitro at the 3'-end, contains [3'-(3)H]poly(A); when labeled in vivo with [(3)H]A, it contains [(3)H](Ap)(n)A(OH). The data establish that... YpGpGp(Ap)([unk])A(OH) is the 3'-terminal sequence of poliovirus RNA, Type 1 (Mahoney). Since this mammalian virus reproduces in the cell cytoplasm, these observations may modify prior interpretations of the function of polyadenylate ends on messenger RNAs.
MeSH Terms
Adenine Nucleotides/analysis
Adenosine
Base Sequence
Borohydrides
Centrifugation, Density Gradient
Chemical Phenomena
Chemistry
Chromatography, Ion Exchange
Coliphages/enzymology
Electrophoresis
Hydrolysis
Hydroxides
Isotope Labeling
Micropore Filters
Nucleotides/analysis
Oxidation-Reduction
Pancreas/enzymology
Periodic Acid
Phosphoric Diester Hydrolases
Phosphorus Isotopes
Poliovirus/analysis
Polynucleotides/analysis
RNA, Viral/analysis
Ribonucleases
Sodium Dodecyl Sulfate
Tritium
Chemicals
Adenine Nucleotides
Borohydrides
Hydroxides
Nucleotides
Phosphorus Isotopes
Polynucleotides
RNA, Viral
Tritium
Periodic Acid
Sodium Dodecyl Sulfate
Ribonucleases
Phosphoric Diester Hydrolases
Adenosine
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Yogo Y
Wimmer E
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