Favre A, Moreno G, Blondel M O, Kliber J, Vinzens F, Salet C
Biochem Biophys Res Commun. 1986 Dec 15;141(2):847-54. doi: 10.1016/s0006-291x(86)80250-9.
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.
在含有0.1 mM 4-硫尿苷（S4U）的条件下培养的猴肾细胞（CV-1），随后在365 nm波长下光照，会表现出明显的RNA合成抑制。4小时的S4U孵育和45 KJ/m²的剂量可获得最大效应（约40%）。在这些条件下，在酚-氯仿提取过程中，高达20%的总细胞RNA保留在间期。与蛋白质交联的RNA部分占S4U掺入过程中合成的3H-尿苷标记RNA的50%，而对照样品则小于10%。这有力地表明，掺入RNA链中的S4U充当光亲和探针。上述数据为在非破坏性条件下研究体内RNA-蛋白质相互作用的方法提供了基础。
