Functions of early (AP-2) and late (AIP1/ALIX) endocytic proteins in equine infectious anemia virus budding

J Biol Chem. 2005 Dec 9;280(49):40474-80. doi: 10.1074/jbc.M509317200. Epub 2005 Oct 7.

Abstract

The proline-rich L domains of human immunodeficiency virus 1 (HIV-1) and other retroviruses interact with late endocytic proteins during virion assembly and budding. In contrast, the YPDL L domain of equine infectious anemia virus (EIAV) is apparently unique in its reported ability to interact both with the mu2 subunit of the AP-2 adaptor protein complex and with ALG-2-interacting protein 1 (AIP1/Alix) protein factors involved in early and late endosome formation, respectively. To define further the mechanisms by which EIAV adapts vesicle trafficking machinery to facilitate virion production, we have examined the specificity of EIAV p9 binding to endocytic factors and the effects on virion production of alterations in early and late endocytic protein expression. The results of these studies demonstrated that (i) an approximately 300-residue region of AIP1/Alix-(409-715) was sufficient for binding to the EIAV YPDL motif; (ii) overexpression of AIP1/Alix or AP-2 mu2 subunit specifically inhibited YPDL-mediated EIAV budding; (iii) virion budding from a replication-competent EIAV variant with its L domain replaced by the HIV PTAP sequence was inhibited by wild type or mutant mu2 to a level similar to that observed when a dominant-negative mutant of Tsg101 was expressed; and (iv) overexpression or siRNA silencing of AIP1/Alix and AP-2 revealed additive suppression of YPDL-mediated EIAV budding. Taken together, these results indicated that both early and late endocytic proteins facilitate EIAV production mediated by either YPDL or PTAP L domains, suggesting a comprehensive involvement of endocytic factors in retroviral assembly and budding that can be accessed by distinct L domain specificities.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Protein Complex 2 / genetics
  • Adaptor Protein Complex 2 / physiology*
  • Animals
  • COS Cells
  • Carrier Proteins / physiology
  • Chlorocebus aethiops
  • DNA, Viral / genetics
  • Endocytosis*
  • Gene Silencing
  • Glutathione Transferase / genetics
  • HeLa Cells
  • Humans
  • Infectious Anemia Virus, Equine / genetics
  • Infectious Anemia Virus, Equine / physiology*
  • Mutagenesis
  • Protein Subunits / genetics
  • Protein Subunits / physiology
  • Proteins / genetics
  • Proteins / physiology*
  • Proviruses / genetics
  • RNA, Small Interfering
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae
  • Transfection
  • Two-Hybrid System Techniques
  • Viral Proteins / physiology

Substances

  • Adaptor Protein Complex 2
  • Carrier Proteins
  • DNA, Viral
  • Protein Subunits
  • Proteins
  • RNA, Small Interfering
  • Recombinant Fusion Proteins
  • Viral Proteins
  • Glutathione Transferase