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Immunological studies of RNA polymerase II using antibodies to subunits of Drosophila and wheat germ enzyme.

Publication ,  Journal Article
Weeks, JR; Coulter, DE; Greenleaf, AL
Published in: J Biol Chem
May 25, 1982

We induced goat antibodies to Drosophila RNA polymerase II and rabbit antibodies to the isolated 215,000-dalton and 140,000-dalton polymerase II subunits (P215 and P140, respectively). Similarly, we induced rabbit antibodies to wheat germ RNA polymerase II and to the 220,000-dalton subunit and 140,000-dalton subunit (P220 and P140, respectively). Anti-polymerase antibodies precipitated the homologous native enzyme and inhibited its activity in vitro, while several of the anti-subunit sera did neither. The anti-Drosophila P215 serum specifically labeled RNA polymerase II fixed in situ on polytene chromosomes. We reacted the antibodies with polymerase subunits separated by sodium dodecyl sulfate gel electrophoresis and electrophoretically transferred to nitrocellulose ("protein blotting"). Each antibody to whole polymerase reacted with multiple subunits, while the anti-subunit sera each reacted specifically with the subunit employed as immunogen. The anti-subunit sera also cross-reacted with the analogous subunit from several heterologous polymerases II (from yeast, wheat germ, Drosophila, and calf thymus), demonstrating shared subunit-specific determinants in polymerase II from widely divergent organisms. The anti-polymerase sera also showed cross-reactivity with subunits of heterologous enzymes, but only in one case did the cross-reactivity involve subunits other than the two largest ones. Specifically, the goat anti-Drosophila polymerase serum displayed easily detectable cross-reactivity with four low molecular weight subunits of calf thymus polymerase II, providing a unique demonstration of antigenic relatedness of small RNA polymerase II subunits from different higher eukaryotes.

Duke Scholars

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

May 25, 1982

Volume

257

Issue

10

Start / End Page

5884 / 5892

Location

United States

Related Subject Headings

  • Triticum
  • Thymus Gland
  • RNA Polymerase II
  • Plants
  • Macromolecular Substances
  • Immunodiffusion
  • Fluorescent Antibody Technique
  • Epitopes
  • Drosophila
  • DNA-Directed RNA Polymerases
 

Citation

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Weeks, J. R., Coulter, D. E., & Greenleaf, A. L. (1982). Immunological studies of RNA polymerase II using antibodies to subunits of Drosophila and wheat germ enzyme. J Biol Chem, 257(10), 5884–5892.
Weeks, J. R., D. E. Coulter, and A. L. Greenleaf. “Immunological studies of RNA polymerase II using antibodies to subunits of Drosophila and wheat germ enzyme.J Biol Chem 257, no. 10 (May 25, 1982): 5884–92.
Weeks JR, Coulter DE, Greenleaf AL. Immunological studies of RNA polymerase II using antibodies to subunits of Drosophila and wheat germ enzyme. J Biol Chem. 1982 May 25;257(10):5884–92.
Weeks, J. R., et al. “Immunological studies of RNA polymerase II using antibodies to subunits of Drosophila and wheat germ enzyme.J Biol Chem, vol. 257, no. 10, May 1982, pp. 5884–92.
Weeks JR, Coulter DE, Greenleaf AL. Immunological studies of RNA polymerase II using antibodies to subunits of Drosophila and wheat germ enzyme. J Biol Chem. 1982 May 25;257(10):5884–5892.

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

May 25, 1982

Volume

257

Issue

10

Start / End Page

5884 / 5892

Location

United States

Related Subject Headings

  • Triticum
  • Thymus Gland
  • RNA Polymerase II
  • Plants
  • Macromolecular Substances
  • Immunodiffusion
  • Fluorescent Antibody Technique
  • Epitopes
  • Drosophila
  • DNA-Directed RNA Polymerases