title: A Nonradioactive Assay to Measure Production and Processing of Ribosomal RNA by 4sU-Tagging.
abstract: In vivo metabolic pulse labeling is a classical approach to assess production and processing of ribosomal RNA (rRNA). However, conventional labeling techniques can be indirect and require work with radioactivity. Here, we describe in detail a protocol for in vivo metabolic labeling, purification, and readout of nascent rRNA by 4-thiouridine (4sU). We propose 4sU labeling as standard nonradioactive technique for the analysis of rRNA metabolism during ribosome biogenesis.
authors: Sir William Dunn School of Pathology, University of Oxford, South Parks Road, OX1 3RE, Oxford, UK.
authors: Kaspar
authors: K
authors: Burger
authors: Department of Molecular Epigenetics, Helmholtz Center Munich and Center for Integrated Protein Science Munich (CIPSM), Marchioninistrasse 25, 81377, Munich, Germany. eick@helmholtz-muenchen.de.
authors: Dirk
authors: D
authors: Eick
mesh_terms: D001681:Biological Assay; D002460:Cell Line; D057930:Click Chemistry; D006801:Humans; D012323:RNA Processing, Post-Transcriptional; D012335:RNA, Ribosomal / Q000235:genetics*; D012270:Ribosomes / Q000235:genetics / Q000378:metabolism; D013194:Staining and Labeling; D013891:Thiouridine; D014158:Transcription, Genetic
chemical_list: D012335:RNA, Ribosomal; D013891:Thiouridine
keywords: 4-thiouridine; Click chemistry; Metabolic pulse labeling; Nascent RNA; Ribosome biogenesis; rRNA synthesis