Immunoprecipitation of DNA-protein complexes cross-linked by cis-diamminedichloroplatinum.
For the study of in vitro and in vivo DNA-protein interactions, cross-linking reactions driven by UV or formaldehyde have been frequently used, followed by standard protocols of immunoprecipitation and analysis of the DNA isolated from the complexes. Here we present a basically modified method to analyze the DNA-protein cross-linked complexes obtained by an alternative cross-linking reagent. The innovations presented here include cross-linking by cis-diamminedichloroplatinum II, a fast method to isolate DNA-protein complexes using gel-filtration chromatography, and a modified procedure to obtain specific immunocomplexes that can be analyzed either for DNA or for protein content. The application of this method to two nuclear proteins from chicken liver nuclei is described.
Chichiarelli
Silvia
S
Istituto Pasteur-Fondazione Cenci Bolognetti, Università 'La Sapienza', Rome 00185, Italy.
Coppari
Sabina
S
Turano
Carlo
C
Eufemi
Margherita
M
Altieri
Fabio
F
Ferraro
Anna
A
D000199:Actins / Q000032:analysis* / Q000378:metabolism; D000818:Animals; D002645:Chickens; D002945:Cisplatin / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000737:chemistry; D004247:DNA / Q000032:analysis* / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000032:analysis* / Q000378:metabolism; D007535:Isomerases / Q000032:analysis* / Q000378:metabolism; D046911:Macromolecular Substances; D009687:Nuclear Proteins / Q000032:analysis / Q000378:metabolism; D011233:Precipitin Tests / Q000379:methods*
D000199:Actins; D003432:Cross-Linking Reagents; D006360:Heat-Shock Proteins; D046911:Macromolecular Substances; D009687:Nuclear Proteins; D004247:DNA; D007535:Isomerases; D002945:Cisplatin