Tsg101-gag interaction and use thereof

Abstract

Isolated protein complexes are provided comprising Tsg101 and HIV GAG or GAGp6. The protein complexes are useful in screening assays for selecting compounds effective in modulating the Tsg101-HIV GAG or GAGp6 interaction within the protein complexes.

1 . A method for inhibiting HIV viral budding from a host cell, comprising reducing the concentration of a protein complex in the cell, said protein complex having a first protein which is Tsg101 interacting with a second protein which is HIV GAG. 
     
     
         2 . The method of  claim 1 , wherein said reducing step comprising interfering with an interaction between said first protein and said second protein. 
     
     
         3 . The method of  claim 2 , wherein said reducing step comprises administering to the cell a compound capable of interfering with an interaction between said first protein and said second protein. 
     
     
         4 . The method of  claim 3 , wherein said compound is capable of binding Tsg101. 
     
     
         5 . The method of  claim 3 , wherein said compound is capable of binding the UEV domain of Tsg101 protein. 
     
     
         6 . The method of  claim 5 , wherein said compound is an antibody immunoreactive with Tsg101. 
     
     
         7 . The method of  claim 5 , wherein said compound is a nucleic acid encoding an antibody immunoreactive with Tsg101. 
     
     
         8 . The method of  claim 7 , wherein said antibody is a single-chain antibody. 
     
     
         9 . The method of  claim 5 , wherein said compound is a peptide having a contiguous span of from 7 to 50 amino acid residues of HIV GAG, said contiguous span encompassing the late domain motif. 
     
     
         10 . The method of  claim 1 , wherein said reducing step comprises reducing the concentration of Tsg101 in the cell. 
     
     
         11 . The method of  claim 10 , wherein said step of reducing the concentration of Tsg101 in the cell comprises administering to the cell a nucleic acid molecule that induce the degradation of RNA transcripts encoding Tsg101. 
     
     
         12 . The method of  claim 11 , wherein said nucleic acid molecule is an antisense compound specifically hybridizing to a Tsg101 nucleic acid. 
     
     
         13 . The method of  claim 11 , wherein said nucleic acid molecule is a ribozyme compound specifically hybridizing to a Tsg101 nucleic acid. 
     
     
         14 . The method  claim 11 , wherein said nucleic acid molecule is an siRNA or an expression vector expressing an shRNA. 
     
     
         15 . The method of  claim 13 , wherein said expression vector comprises a promoter operably linked to an shRNA-encoding nucleic acid. 
     
     
         16 . An expression vector comprising a promoter operably linked to a nucleic acid encoding an shRNA capable of inducing degradation of Tsg101 transcript. 
     
     
         17 . A host cell comprising the expression vector of  claim 16 . 
     
     
         18 . An expression vector comprising a promoter operably linked to a nucleic acid encoding an shRNA capable of hybridizing to a region of an HIV transcript encoding the GAG polypeptide, said region encoding HIV GAGp6. 
     
     
         19 . A host cell comprising the expression vector of  claim 18 .