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Oligomeric properties of adeno-associated virus Rep68 reflect its multifunctionality

Francisco Zarate-Perez1, Jorge Mansilla-Soto, Martino Bardelli

  • 1Department of Physiology and Biophysics, Virginia Commonwealth University School of Medicine, Richmond, VA, USA.

Journal of Virology
|November 16, 2012
PubMed

Insights

The adeno-associated virus Rep68 protein

Area of Science:

  • Molecular biology
  • Virology
  • Biochemistry

Background:

  • Adeno-associated virus (AAV) Rep proteins, Rep68 and Rep78, are crucial for viral replication, transcription, and integration.
  • Rep proteins possess a helicase domain (SF3 family) and a DNA-binding domain, contributing to their multifunctionality.
  • The oligomeric state of Rep68 is poorly understood due to aggregation issues.

Purpose of the Study:

  • To elucidate the oligomeric character of the AAV Rep68 protein.
  • To investigate the factors influencing Rep68's oligomerization and its dynamic behavior.

Main Methods:

  • Sedimentation velocity ultracentrifugation
  • Electron microscopy
  • Hydrodynamic modeling
  • In vitro aggregation studies

Main Results:

  • Residue Cys151 was identified as inducing Rep68 aggregation in vitro.
  • Rep68 exhibits concentration-dependent dynamic oligomerization, forming monomers, dimers, and ring structures of 7-8 members.
  • ATP or ADP binding promotes the formation of larger, stacked ring complexes.

Conclusions:

  • Rep68 displays flexible oligomeric behavior essential for its diverse roles in the AAV life cycle.
  • Understanding Rep68's oligomerization provides insights into AAV replication and gene regulation mechanisms.

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