Structural studies on yeast RNA polymerases. Existence of common subunits in RNA polymerases A(I) and B(II).
The subunits of yeast RNA polymerases A(I) and B(II) were characterized using several techniques. The present studies demonstrate that the A and B enzymes possess three subunits, which are indistinguishable on the basis of molecular weight, isoelectric point, and fingerprint pattern. The three common subunits belong to the small molecular weight components of the enzymes. By polyacrylamide gel electrophoresis with sodium dodecyl sulfate they migrate with apparent molecular weights of 27,000, 23,000, and 14,500, respectively. A two-dimensional subunit mapping technique on polyacrylamide gel was used to separate the subunits according to isoelectric point and molecular weight. The common polypeptides co-migrated on three spots corresponding to isoelectric points of 9.2 (27,000), 4.5 (23,000), and 4.6 (14,500). The fingerprints of the 35S-labeled tryptic peptides of the presumptive common subunits were found to be essentially identical. Finally, the presence of common subunits was supported by the fact that antibodies against pure RNA polymerase A cross-react with and inhibit RNA polymerase B. Except for the common subunits, it is likely that RNA polymerases A and B are primarily made of distinct gene products for the following reasons. A total of 13 polypeptide chains are present in enzyme A, whereas 10 polypeptides are found in enzyme B. The molecular weight, isoelectric point, and sulfur content of the majority of these polypeptide chains are different in the two enzymes. No similarity was found in the 35S-peptide fingerprint from a number of A and B subunits of slightly different molecular weight. Finally, antibodies against the largest subunit from RNA polymerase A do not cross-react with or inhibit RNA polymerase B. The data are discussed in terms of structural organization of eukaryotic RNA polymerases.
D012321:DNA-Directed RNA Polymerases / Q000276:immunology / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000276:immunology / Q000378:metabolism; D046911:Macromolecular Substances; D010446:Peptide Fragments / Q000032:analysis; D011233:Precipitin Tests; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013045:Species Specificity
D007527:Isoenzymes; D046911:Macromolecular Substances; D010446:Peptide Fragments; D012321:DNA-Directed RNA Polymerases