An assay for studying ubiquitylation of RNA polymerase II and other proteins in crude yeast extracts.
In recent years, much research effort has been invested in understanding the role and importance of protein ubiquitylation. This chapter presents protocols that enable ubiquitylation of RNAPII and other proteins to be reconstituted in crude yeast extracts in vitro. These procedures, combined with the ease of genetic manipulations in yeast, provide a new powerful tool for investigating the mechanisms underlying protein ubiquitylation.
Reid
James
J
Cancer Research United Kingdom, London Research Institute, Clare Hall Laboratories, South Mimms, Hertfordshire.
Svejstrup
Jesper Q
JQ
D001681:Biological Assay / Q000379:methods*; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D012319:RNA Polymerase II / Q000302:isolation & purification / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000302:isolation & purification / Q000378:metabolism*; D025801:Ubiquitin / Q000235:genetics / Q000378:metabolism*
D002868:Chromosomal Proteins, Non-Histone; C451742:DEF1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D025801:Ubiquitin; D012319:RNA Polymerase II