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RNA polymerase activity of Amsacta moorei entomopox virions
Abstract:
Optimum conditions for in vitro RNA synthesis by the entomopoxvirus from Amsacta moorei, except for a temperature optimum of 26 degrees, were similar to those reported for vaccinia. Incorporation of 3H-ATP in the presence of CTP, GTP and UTP was significantly inhibited by actinomycin D; incorporation of 3H-ATP alone was not. The products formed by incorporation of 3H-ATP alone or with the three other nucleotides contained polyadenylic acid sequences. The sedimentation coefficient of the 3H-ATP-labeled product formed in the presence of CTP, GTP and UTP was reduced from 8-23S to 3-5S after RNase treatment. The product formed from 3H-ATP alone had an apparent sedimentation value of 3-5S.
Insights
This study investigates in vitro RNA synthesis in Amsacta moorei entomopoxvirus, finding polyadenylic acid sequences in RNA products. Optimal conditions resemble vaccinia virus, with actinomycin D inhibiting RNA synthesis.
Area of Science:
- Virology
- Molecular Biology
- Biochemistry
Background:
- Entomopoxviruses are large DNA viruses that infect insects.
- Understanding their RNA synthesis is crucial for viral replication mechanisms.
- Amsacta moorei entomopoxvirus (AmEPV) serves as a model for studying these viruses.
Purpose of the Study:
- To characterize the in vitro RNA synthesis of AmEPV.
- To determine the optimal conditions for RNA synthesis.
- To analyze the nature and properties of the synthesized RNA products.
Main Methods:
- In vitro RNA synthesis assays using AmEPV extracts.
- Optimization of reaction conditions including temperature and nucleotide incorporation.
- Analysis of RNA products using sedimentation coefficients and RNase treatment.
- Investigation of actinomycin D inhibition.
Main Results:
- Optimal in vitro RNA synthesis occurred at 26°C, similar to vaccinia virus.
- Actinomycin D significantly inhibited the incorporation of 3H-ATP when other nucleotides were present.
- Synthesized RNA products, whether with or without other nucleotides, contained polyadenylic acid sequences.
- RNase treatment reduced the sedimentation coefficient of the full RNA product from 8-23S to 3-5S, while the product from 3H-ATP alone showed a 3-5S value.
Conclusions:
- AmEPV possesses an RNA synthesis system with characteristics similar to vaccinia virus.
- The virus produces polyadenylic acid sequences, suggesting a role in RNA processing or stability.
- Actinomycin D's differential effect indicates specific mechanisms in RNA synthesis pathways.