Cross-linking of proteins to DNA in newly synthesized chromatin by diisopropylfluorophosphate, a serine protease inhibitor
1984; Elsevier BV; Volume: 123; Issue: 1 Linguagem: Inglês
10.1016/0006-291x(84)90408-x
ISSN1090-2104
AutoresKen‐Ichiro Tsutsui, Koji Aoyama, Takuzo Oda,
Tópico(s)RNA Interference and Gene Delivery
ResumoWhen nascent DNA of SV40 pulse labeled with [α-32P]dCTP in a permeable cell system was treated in situ with diisopropylfluorophosphate (DFP), a significant fraction of radioactivity was found to be covalently complexed with proteins. The adduct formation was demonstrated by density separation in CsCl, selective precipitation of the complexed DNA with SDS-KCl, and visualization of cross-linked proteins after SDS-PAGE. No cross-linking occurred with mature SV40 chromatin labeled in vivo and extracted from nuclei of infected cells. The DFP-induced DNA-protein cross-linking reaction appears to involve the protein's sulfhydryl groups since pretreatment with some sulfhydryl reagents completely inhibited the reaction.
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