Compositions and methods using eukaryotic Rad5

Abstract

The invention relates to complexes of eukaryotic Rad52 protein and nucleic acids, and methods of using the complexes.

1 . A composition comprising a complex of a first single stranded nucleic acid and isolated Rad52 protein from a higher eukaryote.  
     
     
         2 . A composition according to  claim 1  wherein said complex is capable of mediating the annealing of said first nucleic acid to a complementary second single stranded nucleic acid.  
     
     
         3 . A composition according to  claim 1  wherein said Rad52 protein is a mammalian Rad52 protein.  
     
     
         4 . A composition according to  claim 1  wherein said Rad52 protein is a human Rad52 protein.  
     
     
         5 . A composition according to  claim 1  further comprising a second single stranded nucleic acid complexed with isolated Rad52 protein from a higher eukaryote.  
     
     
         6 . A composition according to  claim 5  wherein said second nucleic acid is complementary to said first nucleic acid.  
     
     
         7 . A composition according to  claim 1  further comprising a double stranded nucleic acid comprising second and third single stranded nucleic acids, wherein both said first and said third nucleic acids are complementary to said second nucleic acid.  
     
     
         8 . A method of making double stranded nucleic acid comprising contacting: 
 a) a first single stranded nucleic acid;    b) a second single stranded nucleic acid, wherein said first and second nucleic acids are complementary; and    c) isolated Rad52 protein from a higher eukaryote; under conditions whereby said Rad52 mediates annealing of said first and second nucleic acids.    
     
     
         9 . A method according to  claim 8 , wherein one or both of said nucleic acids are complexed with said isolated Rad52 protein prior to said contacting.  
     
     
         10 . A method according to  claim 8 , wherein said annealing is done in the absence of Mg t2  and cofactors.  
     
     
         11 . A method of accomplishing strand exchange comprising contacting: 
 a) a first single stranded nucleic acid;    b) a double stranded nucleic acid comprising second and third single stranded nucleic acids, wherein both said first and third nucleic acids are complementary to said second nucleic acid; and    c) isolated Rad52 from a higher eukaryote; under conditions whereby said Rad52 mediates the annealing of said first nucleic acid to said second nucleic acid, such that said third nucleic acid is displaced.    
     
     
         12 . A method according to  claim 11  wherein any or all of said nucleic acids are complexed with said Rad52 prior to said contacting.  
     
     
         13 . A method according to  claim 11 , wherein said annealing is done in the absence of Mg t2  and cofactors.  
     
     
         14 . A method of screening for a bioactive agent involved in homologous recombination comprising: 
 a) contacting: 
 i) a candidate bioactive agent;  
 ii) a first single stranded nucleic acid; and  
 iii) isolated Rad52 protein from a higher eukaryote; and  
   b) screening for binding of said candidate and said Rad52 to said nucleic acid.    
     
     
         15 . A method according to  claim 14  wherein said first nucleic acid and said isolated Rad52 are complexed prior to the addition of said candidate agent.  
     
     
         16 . A method of screening for a bioactive agent involved in homologous recombination comprising: 
 a) adding: 
 i) a candidate bioactive agent;  
 ii) a first single stranded nucleic acid; and  
 iii) isolated Rad52 protein from a higher eukaryote to form a mixture; and  
   b) screening said mixture for altered biological activity, when compared to the biological activity of said composition in the absence of said candidate.    
     
     
         17 . A method according to  claim 16  wherein said first nucleic acid and said isolated Rad52 are complexed prior to the addition of said candidate agent.