Copurification of casein kinase II with transcription factor ATF/E4TF3

Tadashi Wada, Toshiyuki Takagi, Yuki Yamaguchi, Hiroyuki Kawase, Masaki Hiramoto, Anwarul Ferdous, Makoto Takayama, Kevin A W Lee, Helen C. Hurst, Hiroshi Handa

Research output: Contribution to journalArticle

28 Scopus citations

Abstract

We have developed a simple method to purify sequence-specific DNA-binding proteins directly from crude cell extracts by using DNA affinity latex beads. The method enabled us to purify not only DNA-binding proteins, but also their associated proteins. Using beads bearing the ATF/E4TF3 site from the adenovirus E4 gene promoter, a protein kinase activity was copurified with the ATF/E4TF3 family. We found that the kinase interacted with ATF1 in vitro efficiently. The kinase did not bind directly to DNA. The kinase mainly phosphorylated ATF1 on serine 36, which was one of target amino acids for casein kinase (CK) II. Biological features of the kinase were the same as those of CKII and an anti-CKII serum reacted with the kinase, indicating that the kinase was CKII. Moreover, it was clearly shown that one of CKII subunits, the CKII a protein bound to glutathione-S-transferase (GST) fusion ATF1 but not GST in vitro. It has been reported that a specific CKII inhibitor, 5,6-dichloro-1-β-D-ribofuranosylbenzimidazole (DRB) inhibits transcription by RNA polymerase II. Taken together, these results suggest that ATF/E4TF3 may recruit the CKII activity to a transcription initiation machinery and stimulate transcription.

Original languageEnglish (US)
Pages (from-to)876-884
Number of pages9
JournalNucleic acids research
Volume24
Issue number5
DOIs
StatePublished - 1996

ASJC Scopus subject areas

  • Genetics

Fingerprint Dive into the research topics of 'Copurification of casein kinase II with transcription factor ATF/E4TF3'. Together they form a unique fingerprint.

  • Cite this

    Wada, T., Takagi, T., Yamaguchi, Y., Kawase, H., Hiramoto, M., Ferdous, A., Takayama, M., Lee, K. A. W., Hurst, H. C., & Handa, H. (1996). Copurification of casein kinase II with transcription factor ATF/E4TF3. Nucleic acids research, 24(5), 876-884. https://doi.org/10.1093/nar/24.5.876