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J Biol Chem, Vol. 273, Issue 18, 10827-10830, May 1, 1998
From the McArdle Laboratory for Cancer Research, University of
Wisconsin, Madison, Wisconsin 53706
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 
-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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