title: Rpb3, stoichiometry and sequence determinants of the assembly into yeast RNA polymerase II in vivo.
abstract: Stoichiometry of the third largest subunit (Rpb3) of the yeast RNA polymerase II is a subject of continuing controversy. In this work we utilized immunoaffinity and nickel-chelate chromatographic techniques to isolate the RNA polymerase II species assembled in vivo in the presence of the His6-tagged and untagged Rpb3. The distribution pattern of tagged and untagged subunits among the RNA polymerase II molecules is consistent with a stoichiometry of 1 Rpb3 polypeptide per molecule of RNA polymerase. Deletion of either alpha-homology region (amino acids 29-55 or 226-267) from the Rpb3 sequence abolished its ability to assemble into RNA polymerase II in vivo.
authors.affiliation: McArdle Laboratory for Cancer Research, University of Wisconsin, Madison, Wisconsin 53706, USA.
authors.forename: V
authors.initials: V
authors.lastname: Svetlov
authors.forename: K
authors.initials: K
authors.lastname: Nolan
authors.forename: R R
authors.initials: RR
authors.lastname: Burgess
mesh_terms: D000595:Amino Acid Sequence; D002846:Chromatography, Affinity; D008969:Molecular Sequence Data; D009154:Mutation; D010802:Phylogeny; D012319:RNA Polymerase II / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D012319:RNA Polymerase II; C112110:RPB3 protein, S cerevisiae