Copurification of casein kinase II with transcription factor ATF/E4TF3.
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 alpha 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-beta-D-ribo-furanosylbenzimidazole (DRB) inhibits transcription by RNA polymerase II [Zandomeni et al., (1986) J. Biol. Chem. 261, 3414-3419]. Taken together, these results suggest that ATF/E4TF3 may recruit the CKII activity to a transcription initiation machinery and stimulate transcription.
Wada
T
T
Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Kanagawa, Japan.
Takagi
T
T
Yamaguchi
Y
Y
Kawase
H
H
Hiramoto
M
M
Ferdous
A
A
Takayama
M
M
Lee
K A
KA
Hurst
H C
HC
Handa
H
H
D051696:Activating Transcription Factors; D001798:Blood Proteins / Q000302:isolation & purification* / Q000378:metabolism; D047390:Casein Kinase II; D002846:Chromatography, Affinity / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting; D017346:Protein Serine-Threonine Kinases / Q000302:isolation & purification* / Q000378:metabolism; D014157:Transcription Factors / Q000302:isolation & purification* / Q000378:metabolism
D051696:Activating Transcription Factors; D001798:Blood Proteins; D014157:Transcription Factors; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases