Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Tobacco mosaic virus replicase and replicative structures.

N Young1, J Forney, M Zaitlin

  • 1Department of Plant Pathology, Cornell University, Ithaca, New York 14853.

Journal of Cell Science. Supplement
|January 1, 1987
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Threshold cost of delivering preventive treatment for household contacts of people with multidrug-resistant TB.

The international journal of tuberculosis and lung disease : the official journal of the International Union against Tuberculosis and Lung Disease·2026
Same author

Analysis of Public Comments on Experimental Regulations for Protecting Black Bass during The Spawning Period in Eastern Ontario Reveals Both Stakeholder Acceptance and Skepticism.

Environmental management·2024
Same author

Confronting a post-pandemic new-normal-threats and opportunities to trust-based relationships in natural resource science and management.

Journal of environmental management·2023
Same author

COVID-19 restrictions and recreational fisheries in Ontario, Canada: Preliminary insights from an online angler survey.

Fisheries research·2022
Same author

Real-time surveillance of severe acute respiratory infections in Scottish hospitals: an electronic register-based approach, 2017-2022.

Public health·2022
Same author

Drivers of pro-environmental behaviours among outdoor recreationists: The case of a recreational fishery in Western Canada.

Journal of environmental management·2021

Researchers identified the tobacco mosaic virus (TMV) replicase and characterized its RNA replication products. Studies indicate TMV RNA synthesis occurs in the cytoplasm, not the nucleus, challenging previous assumptions.

Area of Science:

  • Molecular Virology
  • Plant Pathology
  • Biochemistry

Background:

  • Investigating the RNA-dependent RNA polymerase (replicase) responsible for tobacco mosaic virus (TMV) replication has been a long-standing challenge.
  • Previous research identified cell-free enzyme preparations capable of RNA synthesis but lacking template specificity.
  • The role of host-encoded RNA-dependent RNA polymerase (RdRp) in TMV replication remains debated, with general consensus excluding it as the viral replicase.

Purpose of the Study:

  • To characterize the TMV replicase and its synthesized RNA products in vitro.
  • To investigate the intracellular site of TMV RNA replication.
  • To clarify the nature of TMV replicative structures and the polarity of nascent RNA synthesis.

Main Methods:

  • Partially purified enzyme preparations from TMV-infected tobacco tissue were used for in vitro RNA synthesis.

Related Experiment Videos

  • Radiolabeled RNA products were analyzed using agarose gel electrophoresis and hybridization with M13 clones of TMV genome.
  • Solubilization of membrane-bound replicase with CHAPS detergent and purification via Sepharose 4B chromatography were employed.
  • Subcellular fractionation of infected leaves and protoplasts was performed to determine the site of TMV RNA synthesis.

    • Main Results:

    • Fractions exhibiting electrophoretic migration and nuclease sensitivities of replicative form (RF) and replicative intermediate (RI) were isolated.
    • Nascent RNA synthesis in RI-like molecules was restricted to the plus viral strand; RF-like molecules showed synthesis of both plus and minus strands.
    • Solubilized replication complexes were purified, yielding active enzyme preparations.
    • Double-stranded TMV RNA was concentrated in the post-nuclear supernatant, with minimal presence in purified nuclei, indicating extranuclear replication.

    Conclusions:

    • The study provides insights into the biochemical properties of the TMV replicase and the nature of its replication intermediates.
    • Evidence strongly suggests that TMV RNA replication occurs in the cytoplasm, refuting previous suggestions of a nuclear site.
    • The 126,000 Mr TMV protein and replicase activity were found distributed throughout homogenates, complicating earlier localization studies.