Rpb3, stoichiometry and sequence determinants of the assembly into yeast RNA polymerase II in vivo
- PMID: 9556554
- DOI: 10.1074/jbc.273.18.10827
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.
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