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Efficient Recombinant Parvovirus Production with the Help of Adenovirus-derived Systems

Published on: April 23, 2012

Internal polyadenylation of parvoviral precursor mRNA limits progeny virus production

Qinfeng Huang1, Xuefeng Deng, Sonja M Best

  • 1Department of Microbiology, Molecular Genetics and Immunology, University of Kansas Medical Center, Kansas City, Kansas 66160, USA.

Virology
|February 25, 2012
PubMed

Insights

Aleutian Mink Disease Virus (AMDV) limits capsid protein expression by controlling mRNA processing. Inhibiting internal polyadenylation boosts capsid production, viral DNA replication, and progeny virus yield.

Area of Science:

  • Virology
  • Molecular Biology
  • Immunology

Background:

  • Aleutian Mink Disease Virus (AMDV), an Amdovirus, causes persistent infections and immune dysfunction in mink.
  • Previous research identified capsid protein cleavage as a factor in AMDV progeny production.

Purpose of the Study:

  • To investigate a second AMDV strategy for limiting capsid protein expression.
  • To characterize the cis-elements controlling proximal polyadenylation in AMDV.
  • To determine the impact of polyadenylation on viral replication and progeny production.

Main Methods:

  • Characterization of cis-elements (upstream element, AAUAAA signal, downstream element) at the proximal polyadenylation site [(pA)p] in an AMDV infectious clone.
  • Mutation of the AAUAAA signal or downstream element to alter polyadenylation efficiency.
  • Quantification of capsid protein VP1/VP2 expression, viral DNA replication, and progeny virus production.

Main Results:

  • Polyadenylation at the (pA)p site is regulated by an upstream element, AAUAAA signal, and downstream element.
  • Mutations decreasing polyadenylation at (pA)p increased VP1/VP2 expression 2-3 fold.
  • Increased capsid production correlated with enhanced AMDV genome replication and progeny virus yield.

Conclusions:

  • AMDV utilizes internal polyadenylation as a mechanism to limit capsid protein expression.
  • Internal polyadenylation acts as a rate-limiting step for parvovirus replication and progeny production.
  • Findings reveal a link between polyadenylation, capsid synthesis, viral replication, and virus yield in AMDV.

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