Journal Article

Purification, Characterization, and cDNA Cloning of ABP-2 (Aryiphorin Gene-Specific Binding Protein-2) That Specifically Binds to the ABP-1-Binding Sequence in the Arylphorin Gene of <i>Sarcophaga peregrina</i>

Nobuaki Adachi, Takeo Kubo and Shunji Natori

in The Journal of Biochemistry

Published on behalf of The Japanese Biochemical Society

Volume 120, issue 6, pages 1239-1246
Published in print December 1996 | ISSN: 0021-924X
Published online December 1996 | e-ISSN: 1756-2651 | DOI: http://dx.doi.org/10.1093/oxfordjournals.jbchem.a021547
Purification, Characterization, and cDNA Cloning of ABP-2 (Aryiphorin Gene-Specific Binding Protein-2) That Specifically Binds to the ABP-1-Binding Sequence in the Arylphorin Gene of Sarcophaga peregrina

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Previously, we demonstrated that ABP-1 (aryiphorm gene-specific binding protein-1), which is suggested to be the transcriptional activator of the aryiphorin gene of Sarcophaga peregrina, is present in NIH-Sape-4 cells, which do not express aryiphorin. As well as ABP-1, these cells were found to contain another protein (ABP-2) that probably binds to the same sequence as that to which ABP-1 binds [ Adachi, N., Kubo, T., and Natori, S. (1993) J. Biochem. 114, 55–60]. We purified ABP-2 from a nuclear extract of NIH-Sape-4 cells and compared its DNA-binding activity with that of ABP-1. Both ABP-1 and ABP-2 were found to bind to the same sequence in the arylphorin gene with the same affinity and stability, but an ABP-2-specific hypersensitive site was detected by DNase I footprinting analysis. Analyses of proteolytic fragments suggested that both ABP-1 and ABP-2 have Zn fingers showing high similarity with that of AEF-1, a transcriptional repressor of the Drosophila melanogaster alcohol dehydrogenase gene that binds to a sequence very similar to that binding ABP-1 and ABP-2. We isolated a candidate cDNA for ABP-2, and the protein encoded contained nine Zn fingers and regions rich in alanine, glutamine, serine/threonine, glycine, histidine, and asparagine.

Keywords: ABP-1; ABP-2; arylphorin gene; Sarcophaga peregrina; transcription factor

Journal Article.  0 words. 

Subjects: Biochemistry

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