HIV capsid assembly-associated compositions and methods

Abstract

A cell-free method for translation and assembly of retroviral, particularly HIV, capsid and capsid intermediates is disclosed. Also disclosed are novel HIV capsid assembly intermediates and novel host proteins which bind to such assembly intermediates. The invention also includes a screening method for compounds that alter retrovirus capsid assembly, and a method of treating HIV using compounds which inhibit the HIV capsid assembly pathway.

1 - 12 . (canceled) 
     
     
         13 . A method of selecting a compound that alters HIV capsid assembly, the method comprising:
 (a.) adding a test compound to a reaction mixture that includes:
 (i) a cell-free translation mixture that includes a cell-free extract, tRNA, ribosomes, amino acids and an energy source, 
 (ii) an mRNA molecule encoding HIV Gag Pr55, 
 (iii) myristoyl coenzyme A, present at a concentration greater than about 0.1 micromolar, 
   (b.) measuring capsid assembly in the presence of said test compound,   (c.) comparing assembly in the absence of said test compound to assembly in the presence of said test compound, and   (d.) selecting the test compound as a compound that alters HIV capsid assembly if assembly measured in the absence of said test compound is different than assembly measured in the presence of said test compound.   
     
     
         14 . The method of  claim 13 , wherein said measuring of capsid assembly includes measuring formation of 10-S, 80-S, 150-S, 500-S, and 750-S assembly intermediates. 
     
     
         15 - 19 . (canceled) 
     
     
         20 . A method of selecting a compound that alters HIV capsid assembly in cells, the method comprising:
 adding a test compound to cells that are forming retroviral capsids,   measuring the quantity and proportion of capsid assembly intermediates formed within the cells in the presence of said test compound by measuring formation of a 10-S assembly intermediate, an 80-S assembly intermediate, a 150-S assembly intermediate, a 500-S assembly intermediate, and a 750-S assembly intermediate,   comparing the quantity and proportion of the 10-S, 80-S, 150-S, 500-S, and 750-S assembly intermediates formed within the cells in the absence of said test compound to said quantity and proportion of the 10-S, 80-S, 150-S, 500-S and 750-S assembly intermediates formed in the cells in the presence of said test compound, and   selecting the test compound as a compound that alters HIV capsid assembly if the quantity or proportion of the assembly intermediates measured in the presence of said test compound is different than the quantity or proportion of the assembly intermediates measured in the absence of said test compound.   
     
     
         21 - 22 . (canceled)