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Purified MotA protein binds the -30 region of a bacteriophage T4 middle-mode promoter and activates transcription in vitro.

Publication ,  Journal Article
Schmidt, RP; Kreuzer, KN
Published in: J Biol Chem
June 5, 1992

The bacteriophage T4-encoded MotA protein is critical for transcription from T4 middle-mode promoters. However, a direct interaction of this protein with a middle-mode promoter has not previously been demonstrated. We have cloned the motA gene and overexpressed the gene product using the T7 expression system. A simple procedure was then developed to purify the MotA protein to homogeneity. Using the purified protein we have demonstrated that MotA protein binds to the -30 region of the middle-mode promoter PuvsY. This promoter has previously been shown to be a necessary component of a T4 replication origin, and thus MotA is also a T4 origin-binding protein. Modified RNA polymerase purified from T4-infected cells was used to establish middle-mode transcription in vitro. Transcription from PuvsY was markedly enhanced by the addition of MotA protein, whether or not the template contained the cytosine modifications characteristic of T4 DNA. However, transcription from PuvsY was apparently independent of the MotA protein when unmodified RNA polymerase from uninfected cells was used.

Duke Scholars

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

June 5, 1992

Volume

267

Issue

16

Start / End Page

11399 / 11407

Location

United States

Related Subject Headings

  • Viral Proteins
  • Transcription, Genetic
  • Transcription Factors
  • T-Phages
  • Promoter Regions, Genetic
  • Molecular Sequence Data
  • Genes, Viral
  • Gene Expression
  • Escherichia coli
  • Electrophoresis, Polyacrylamide Gel
 

Citation

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Schmidt, R. P., and K. N. Kreuzer. “Purified MotA protein binds the -30 region of a bacteriophage T4 middle-mode promoter and activates transcription in vitro.J Biol Chem 267, no. 16 (June 5, 1992): 11399–407.
Schmidt, R. P., and K. N. Kreuzer. “Purified MotA protein binds the -30 region of a bacteriophage T4 middle-mode promoter and activates transcription in vitro.J Biol Chem, vol. 267, no. 16, June 1992, pp. 11399–407.

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

June 5, 1992

Volume

267

Issue

16

Start / End Page

11399 / 11407

Location

United States

Related Subject Headings

  • Viral Proteins
  • Transcription, Genetic
  • Transcription Factors
  • T-Phages
  • Promoter Regions, Genetic
  • Molecular Sequence Data
  • Genes, Viral
  • Gene Expression
  • Escherichia coli
  • Electrophoresis, Polyacrylamide Gel