Characterization of the molecularly cloned murine α-globin transcription factor CP2 Journal Article
| Authors: | Lim, L. C.; Fang, L.; Swendeman, S. L.; Sheffery, M. |
| Article Title: | Characterization of the molecularly cloned murine α-globin transcription factor CP2 |
| Abstract: | We recently cloned human and murine cDNAs that encode CP2, a transcription factor that interacts with the murine α-globin promoter. In this report, we exploited our ability to express CP2 in bacteria and eukaryotic cells to further investigate factor activities in vitro and in vivo. CP2 expressed in bacteria was significantly enriched and used in a series of DNase I footprinting and electrophoretic gel shift assays. The results suggest that CP2 binds a hyphenated recognition sequence motif that spans one DNA helix turn. In addition, the enriched bacterial protein activated transcription of α-globin promoter templates approximately 3- to 4-fold in vitro. We then tested the effect of elevating CP2 levels 2.5- to 5.5-fold in vivo using both transient and stable transformation assays. When a reporter construct comprised of the intact murine α-globin promoter driving the bacterial chloramphenicol acetyltransferase (CAT) gene was introduced into these overexpressing cells, we observed a 3- to 6-fold increase in CAT activity when compared to cells expressing normal levels of CP2. These results define the CP2 factor binding site in more detail and help characterize the activities of the factor in vivo. |
| Keywords: | promoter region; dna-binding proteins; nonhuman; animal cell; mouse; animal; mice; transcription factor; genetic transcription; transcription, genetic; tumor cells, cultured; bacteria (microorganisms); transcription factors; gene expression regulation; molecular cloning; cloning, molecular; gene expression regulation, neoplastic; molecular sequence data; genetic transfection; kinetics; eukaryota; murinae; immunoblotting; base sequence; binding site; binding sites; cell nucleus; dna binding; oligonucleotide; enzyme assay; complementary dna; globins; oligodeoxyribonucleotides; gel electrophoresis; cell extract; cell strain k 562; dna footprinting; promoter regions (genetics); alpha globin; deoxyribonuclease i; leukemia, myeloid, chronic; felis catus; human; priority journal; article; chloramphenicol acetyltransferase; leukemia, experimental; support, non-u.s. gov't; support, u.s. gov't, p.h.s.; erythroleukemia cell |
| Journal Title: | Journal of Biological Chemistry |
| Volume: | 268 |
| Issue: | 24 |
| ISSN: | 0021-9258 |
| Publisher: | American Society for Biochemistry and Molecular Biology |
| Date Published: | 1993-08-25 |
| Start Page: | 18008 |
| End Page: | 18017 |
| Language: | English |
| PUBMED: | 8349681 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | Article -- Export Date: 1 March 2019 -- Source: Scopus |
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