4-Thiouridine photosensitized RNA-protein crosslinking in mammalian cells.
Monkey kidney cells (CV-1) cultivated in the presence of 0.1 mM 4-thiouridine (S4U) and subsequently illuminated at 365 nm exhibit a marked RNA synthesis inhibition. Maximal effect (approximately 40%) was obtained for a 4 h S4U incubation and a 45 KJ/m2 dose. Under these conditions up to 20% of total cellular RNA is retained at the interphase during phenol-chloroform extraction. The fraction of RNA crosslinked to proteins amounts to 50% of the 3H-uridine labeled RNA synthesized during S4U incorporation and less than 10% for the control samples. This strongly suggests that S4U incorporated within the RNA chains acts as a photoaffinity probe. The data above provide the basis of a method for studying in vivo RNA-protein interactions under non destructive conditions.
D000818:Animals; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D003432:Cross-Linking Reagents; D009698:Nucleoproteins; D010777:Photochemistry; D012313:RNA / Q000096:biosynthesis / Q000378:metabolism*; D013891:Thiouridine / Q000378:metabolism*
D003432:Cross-Linking Reagents; D009698:Nucleoproteins; D013891:Thiouridine; D012313:RNA