Abstract
The 5'-terminal nucleotide sequences of human reovirus double-stranded RNA were determined after labeling the RNA with [(32)P]phosphate by polynucleotide kinase. The 5' terminal were labeled to only a limited extent prior to sequential oxidation, beta-elimination, and phosphomonoesterase treatment, indicating that the terminal phosphates were in a modified, blocked configuration. Each genome segment, after removing the blocking group, contained the same two 5'-terminal sequences: GpApUp in one chain and G(*)pCp in the other. G(*)p is a derivative of guanylic acid, probably 2'-O-methyl-Gp, which renders the 5'-terminal sequence resistant to hydrolysis by alkali. The results indicate that the transcription of reovirus double-stranded RNA strats from the 3' end complementary to the G(*)pCp-terminal, resulting in the synthesis of single-stranded mRNA carrying the same 5' sequence as the G(*)pCp-chain. The presence of a modified nucleotide at the 5' terminus of the strand complementary to the mRNA template is a feature common to another double-stranded RNA virus, cytoplasmic polyhedrosis virus.
MeSH Terms
Autoradiography
Base Sequence
Chromatography, Ion Exchange
Chromatography, Paper
Cytosine Nucleotides/analysis
Electrophoresis, Polyacrylamide Gel
Guanine Nucleotides/analysis
Humans
Nucleotides/analysis
Oligonucleotides/analysis
Phosphorus Radioisotopes
Polynucleotides/analysis
RNA, Viral/analysis
Reoviridae/analysis
Ribonucleases
Chemicals
Cytosine Nucleotides
Guanine Nucleotides
Nucleotides
Oligonucleotides
Phosphorus Radioisotopes
Polynucleotides
RNA, Viral
Ribonucleases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Miura K
Watanabe K
Sugiura M
Shatkin A J
References (18)
18 references, click to expand
-
[Structure of the 5'-terminal of high-polymer nucleic acids--analytical method and its application].
Seikagaku. 1967 Aug;39(8):455-62
PMID: 4869021
-
5'-terminal nucleotide sequence of reovirus mRNA synthesized in vitro.
Nat New Biol. 1972 Jan 26;235(56):105-7
PMID: 4501191
-
Transcription of double-stranded RNA in cytoplasmic polyhedrosis virus in vtro.
Virology. 1973 May;53(1):283-6
PMID: 4574875
-
Analysis of the 5'-terminal nucleotide sequences of ribonucleic acids 1. the 5'-termini of Excherichia coli ribosomal RNA.
J Mol Biol. 1967 Jan 28;23(2):135-48
PMID: 5340184
-
Single-stranded RNA synthesis in vitro by the RNA polymerase associated with cytoplasmic polyhedrosis virus containing double-stranded RNA.
J Biochem. 1973 Jul;74(1):117-25
PMID: 4582169
-
RNA polymerase activity in purified reoviruses.
Proc Natl Acad Sci U S A. 1968 Dec;61(4):1462-9
PMID: 4179180
-
Asynchronous synthesis of the complementary strands of the reovirus genome.
Proc Natl Acad Sci U S A. 1971 Feb;68(2):505-8
PMID: 5277109
-
Characterization of a Ribonucleic Acid Polymerase Activity Associated with Purified Cytoplasmic Polyhedrosis Virus of the Silkworm Bombyx mori.
J Virol. 1969 Dec;4(6):857-65
PMID: 16789118
-
5'-Terminal nucleotide sequences of the double-stranded RNA of silkworm cytoplasmic polyhedrosis virus.
J Mol Biol. 1974 Jun 15;86(1):31-48
PMID: 4369455
-
Methylated messenger RNA synthesis in vitro by purified reovirus.
Proc Natl Acad Sci U S A. 1974 Aug;71(8):3204-7
PMID: 4528644
-
Initiation of reovirus mRNA synthesis in vitro.
Nat New Biol. 1971 Apr 7;230(14):169-72
PMID: 4324352
-
Reovirus: RNA polymerase activity in purified virions.
Biochem Biophys Res Commun. 1968 Dec 30;33(6):895-901
PMID: 5750853
-
Separation of ten reovirus genome segments by polyacrylamide gel electrophoresis.
J Virol. 1968 Oct;2(10):986-91
PMID: 5723704
-
The 5'-terminal nucleotides of T7 bacteriophage deoxyribonucleic acid.
J Mol Biol. 1966 Jan;15(1):49-61
PMID: 5912053
-
Segments of genome of viruses containing double-stranded ribonucleic acid.
J Mol Biol. 1970 Jul 28;51(2):247-53
PMID: 5485904
-
Phosphorylation of nucleic acid by an enzyme from T4 bacteriophage-infected Escherichia coli.
Proc Natl Acad Sci U S A. 1965 Jul;54(1):158-65
PMID: 5323016
-
5'-Terminal structure of messenger RNA transcribed by RNA polymerase of silkworm cytoplasmic polyhedrosis virus containing double-stranded RNA.
J Mol Biol. 1974 Jun 15;86(1):21-30
PMID: 4605463
-
Guanosine-5'-diphosphate at the 5' termini of reovirus RNA: evidence for a segmented genome within the virion.
J Mol Biol. 1971 Nov 14;61(3):643-53
PMID: 5167308