Related Experiment Videos
Rhinovirus type 14 RNA polymerase complexes
Abstract:
Rhinovirus type 14 RNA-dependent RNA polymerase complexes were isolated from microsomal and soluble fraction of infected KB cells. Maximum activities were measured at at 6 and 7 hours post inoculation (p.i.) for microsomal and soluble polymerases, respectively. Both polymerase activities are considerably reduced by 8 to 9 hours, p.i., and interval in which the in vivo rate of synthesis of viral RNA is maximal. In vitro RNA products of RNA polymerases in both fractions consist of ribonuclease-sensitive and ribonuclease-resistant RNA of heterogeneous sizes. Detergent treatment of the microsomal RNA polymerase(s) did not affect the total amount of RNA synthesized, the proportion of ribonuclease-sensitive RNA synthesized or the size of the RNA products. The data suggest that RV14RNA polymerase complexes are intially associated with membranes but are then irreversibly released into the soluble phase of the cytoplasm; possible explanations for this phenomena are discussed.
Insights
Rhinovirus type 14 RNA polymerase complexes are initially membrane-associated but later released into the cytoplasm. This study tracks viral RNA synthesis and polymerase activity over time in infected cells.
Area of Science:
- Virology
- Molecular Biology
- Cell Biology
Background:
- Rhinovirus type 14 (RV14) infection involves viral RNA replication.
- Understanding the localization and activity of viral RNA-dependent RNA polymerase is crucial for viral replication studies.
Purpose of the Study:
- To investigate the characteristics and localization of RV14 RNA-dependent RNA polymerase complexes during infection.
- To correlate polymerase activity with viral RNA synthesis rates in infected cells.
Main Methods:
- Isolation of microsomal and soluble RNA polymerase complexes from infected KB cells.
- Measurement of polymerase activities at different time points post-inoculation.
- Analysis of in vitro RNA products, including size heterogeneity and ribonuclease sensitivity.
- Assessment of detergent effects on microsomal RNA polymerase activity.
Main Results:
- Peak polymerase activities were observed at 6-7 hours post-inoculation for microsomal and soluble fractions, respectively.
- Both activities decreased significantly by 8-9 hours post-inoculation, coinciding with maximal in vivo viral RNA synthesis.
- Synthesized RNA products were heterogeneous in size and included both ribonuclease-sensitive and resistant forms.
- Detergent treatment did not alter the RNA synthesis profile of microsomal polymerases.
Conclusions:
- RV14 RNA polymerase complexes appear to transition from a membrane-associated state to a soluble cytoplasmic form.
- This transition may be an irreversible release mechanism.
- The findings provide insights into the dynamic behavior of viral RNA polymerase during replication.