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Involvement of vacuolar proton ATPase in Junin virus multiplication

V Castilla1, L M Palermo, C E Coto

  • 1Laboratorio de Virología, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina.

Archives of Virology
|April 24, 2001
PubMed

Insights

Vacuolar-proton ATPase (V-H+ ATPase) inhibitors like bafilomycin A1 reduce Junin virus (JV) replication by blocking early entry and release. This confirms JV uses the endocytic pathway for cell entry.

Area of Science:

  • Virology
  • Cell Biology
  • Biochemistry

Background:

  • Junin virus (JV) is a significant human pathogen.
  • The role of vacuolar-proton ATPase (V-H+ ATPase) in viral replication is increasingly recognized.
  • Understanding JV entry mechanisms is crucial for therapeutic development.

Purpose of the Study:

  • To investigate the role of V-H+ ATPase in Junin virus replication.
  • To determine the stage of the JV multiplication cycle affected by V-H+ ATPase inhibitors.
  • To confirm the endocytic pathway involvement in JV cell entry.

Main Methods:

  • Treatment of infected Vero and BHK-21 cells with V-H+ ATPase inhibitors (bafilomycin A1, concanamycin A).
  • Assessment of viral protein expression and infectious virus production.
  • Analysis of virus attachment, penetration, and release.
  • Immunofluorescence staining for viral glycoproteins.
  • Correlation of V-H+ ATPase activity with intracellular acidification.

Main Results:

  • Bafilomycin A1 and concanamycin A significantly reduced JV replication and viral protein expression.
  • Inhibitors primarily blocked early-stage JV multiplication, specifically preventing virus penetration but not attachment.
  • A correlation was observed between V-H+ ATPase inhibition, reduced intracellular acidification, and decreased JV yield.
  • Concanamycin A also interfered with the release of infectious JV particles.
  • Intracellular transport of JV glycoproteins to the cell membrane was not affected.

Conclusions:

  • V-H+ ATPase activity is essential for efficient Junin virus replication.
  • JV entry into host cells occurs via the endocytic pathway.
  • Inhibiting V-H+ ATPase can disrupt multiple stages of the JV life cycle, including entry and particle release.

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