Epitope mapping and functional characterization of monoclonal antibodies specific for the α subunit of Escherichia coli RNA polymerase

Karim A. Sharif, Noboyuki Fujita, Ruzhong Jin, Kazuhiko Igarashi, Akira Ishihama, Joseph S. Krakow

研究成果: Article査読

6 被引用数 (Scopus)

抄録

The epitopes have been localized for a set of monoclonal antibodies specific for the α subunit of the Escherichia coli RNA polymerase. The antibodies are classified into three groups based on their epitopic assignments. Group 1, mAb 123C2, maps in the N terminus of α between amino acids 1 and 23; Group 2 antibodies (mAb 129C4, mAb 124D1 and mAb 121C5) map in the central region between amino acids 190 and 210; Group 3 antibodies (mAb 130B1 and mAb 125C6) map in the C terminus between amino acids 310 and 320. mAb 130C2 is anomalous since it maps to the N terminus between amino acids 1 and 23 as well as to the C terminus between amino acids 320 and 329. The antibodies were used to investigate the role of α in transcription activation with cAMP receptor protein-dependent promoters. Three antibodies (130C2, 121C5, and 125C6) inhibited cAMP receptor protein-dependent initiation with lac P+ but not with lac UV5 or gal P+. Inhibition was observed with free RNA polymerase and the closed promoter complex; the preformed open promoter complex was insensitive. Only lac P+ was sensitive to these anti-α antibodies supporting the concept that the mode of interaction of RNA polymerase with cAMP receptor protein differs between lac P+ and gal P+.

本文言語English
ページ(範囲)23655-23660
ページ数6
ジャーナルJournal of Biological Chemistry
269
38
出版ステータスPublished - 1994 9 23
外部発表はい

ASJC Scopus subject areas

  • 生化学
  • 分子生物学
  • 細胞生物学

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