RNA polymerase II from wheat germ: a cross-linking study of subunits topography.
RNA polymerase II purified from wheat germ has been treated with a series of cleavable bifunctional reagents and the resulting crosslinked products have been analyzed by diagonal electrophoresis. The results indicate that the three largest subunits (220, 140, and 42,40 kDa, respectively) form a core around which the smaller subunits are bound. The 220- and 140-kDa subunits can be also crosslinked together in the absence of bifunctional reagents by disulfide bond(s) formation. The 27-, 16.3- and 16-kDa subunits appear to be close to the largest subunit (220 kDa). The 21-kDa subunit is close to the 27- and 25-kDa subunits. The reaction of monofunctional reagents with the enzyme shows that the 42,40-kDa subunit is partially hidden in the interior of the protein molecule. On the basis of these results a model of the quaternary structure of the enzyme is proposed.
Cervoni
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Department of Biochemical Sciences A. Rossi Fanelli, University La Sapienza, Rome, Italy.
Ferraro
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A
Giartosio
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A
Wang
C
C
Turano
C
C
D003432:Cross-Linking Reagents; D004119:Dimethyl Adipimidate; D004120:Dimethyl Suberimidate; D004220:Disulfides / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D046911:Macromolecular Substances; D011487:Protein Conformation; D012319:RNA Polymerase II / Q000737:chemistry*; D013388:Succinimides; D014908:Triticum / Q000201:enzymology*
D003432:Cross-Linking Reagents; D004220:Disulfides; D046911:Macromolecular Substances; D013388:Succinimides; D004119:Dimethyl Adipimidate; D004120:Dimethyl Suberimidate; C035759:3,3'-dithiobis(sulfosuccinimidyl propionate); D012319:RNA Polymerase II