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. 1986 Dec 15;141(2):847-54.
doi: 10.1016/s0006-291x(86)80250-9.

4-Thiouridine photosensitized RNA-protein crosslinking in mammalian cells

A Favre, G Moreno, M O Blondel, J Kliber, F Vinzens, C Salet

4-Thiouridine photosensitized RNA-protein crosslinking in mammalian cells

A Favre et al. Biochem Biophys Res Commun. .

Abstract

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.

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