RNA binding and replication by the poliovirus RNA polymerase
Thesis/Dissertation
·
OSTI ID:6622841
RNA binding and RNA synthesis by the poliovirus RNA-dependent RNA polymerase were studied in vitro using purified polymerase. Templates for binding and RNA synthesis studies were natural RNAs, homopolymeric RNAs, or subgenomic poliovirus-specific RNAs synthesized in vitro from cDNA clones using SP6 or T7 RNA polymerases. The binding of the purified polymerase to poliovirion and other RNAs was studied using a protein-RNA nitrocellulose filter binding assay. A cellular poly(A)-binding protein was found in the viral polymerase preparations, but was easily separated from the polymerase by chromatography on poly(A) Sepharose. The binding of purified polymerase to {sup 32}P-labeled ribohomopolymeric RNAs was examined, and the order of binding observed was poly(G) >>> poly(U) > poly(C) > poly(A). The K{sub a} for polymerase binding to poliovirion RNA and to a full-length negative strand transcript was about 1 {times} 10{sup 9} M{sup {minus}1}. The polymerase binds to a subgenomic RNAs which contain the 3{prime} end of the genome with a K{sub a} similar to that for virion RNA, but binds less well to 18S rRNA, globin mRNA, and subgenomic RNAs which lack portions of the 3{prime} noncoding region.
- Research Organization:
- Florida Univ., Gainesville, FL (USA)
- OSTI ID:
- 6622841
- Country of Publication:
- United States
- Language:
- English
Similar Records
Purification and properties of poliovirus RNA polymerase expressed in Escherichia coli
Poliovirus RNA polymerase: in vitro enzymatic activities, fidelity of replication, and characterization of a temperature-sensitive RNA-negative mutant
Direct measurement of the poliovirus RNA polymerase error frequency in vitro
Journal Article
·
Sat Dec 31 23:00:00 EST 1988
· J. Virol.; (United States)
·
OSTI ID:6204509
Poliovirus RNA polymerase: in vitro enzymatic activities, fidelity of replication, and characterization of a temperature-sensitive RNA-negative mutant
Thesis/Dissertation
·
Mon Dec 31 23:00:00 EST 1984
·
OSTI ID:6941553
Direct measurement of the poliovirus RNA polymerase error frequency in vitro
Journal Article
·
Sun Jan 31 23:00:00 EST 1988
· Journal of Virology; (USA)
·
OSTI ID:6525520
Related Subjects
550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
DAYS LIVING RADIOISOTOPES
DNA REPLICATION
ENZYMES
IN VITRO
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LIGHT NUCLEI
MEMBRANE PROTEINS
MICROORGANISMS
NUCLEI
NUCLEIC ACID REPLICATION
NUCLEIC ACIDS
NUCLEOTIDYLTRANSFERASES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PARASITES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
POLIO VIRUS
POLYMERASES
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RECEPTORS
RNA
RNA POLYMERASES
TRACER TECHNIQUES
TRANSFERASES
VIRUSES
59 BASIC BIOLOGICAL SCIENCES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
DAYS LIVING RADIOISOTOPES
DNA REPLICATION
ENZYMES
IN VITRO
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LIGHT NUCLEI
MEMBRANE PROTEINS
MICROORGANISMS
NUCLEI
NUCLEIC ACID REPLICATION
NUCLEIC ACIDS
NUCLEOTIDYLTRANSFERASES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PARASITES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PHOSPHORUS-GROUP TRANSFERASES
POLIO VIRUS
POLYMERASES
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RECEPTORS
RNA
RNA POLYMERASES
TRACER TECHNIQUES
TRANSFERASES
VIRUSES