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PMID:9422764
| Citation |
Yet, SF, McA'Nulty, MM, Folta, SC, Yen, HW, Yoshizumi, M, Hsieh, CM, Layne, MD, Chin, MT, Wang, H, Perrella, MA, Jain, MK and Lee, ME (1998) Human EZF, a Krüppel-like zinc finger protein, is expressed in vascular endothelial cells and contains transcriptional activation and repression domains. J. Biol. Chem. 273:1026-31 |
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| Abstract |
Members of the erythroid Krüppel-like factor (EKLF) multigene family contain three C-terminal zinc fingers, and they are typically expressed in a limited number of tissues. EKLF, the founding member, transactivates the beta-globin promoter by binding to the CACCC motif. EKLF is essential for expression of the beta-globin gene as demonstrated by gene deletion experiments in mice. Using a DNA probe from the zinc finger region of EKLF, we cloned a cDNA encoding a member of this family from a human vascular endothelial cell cDNA library. Sequence analysis indicated that our clone, hEZF, is the human homologue of the recently reported mouse EZF and GKLF. hEZF is a single-copy gene that maps to chromosome 9q31. By gel mobility shift analysis, purified recombinant hEZF protein bound specifically to a probe containing the CACCC core sequence. In co-transfection experiments, we found that sense but not antisense hEZF decreased the activity of a reporter plasmid containing the CACCC sequence upstream of the thymidine kinase promoter by 6-fold. In contrast, EKLF increased the activity of the reporter plasmid by 3-fold. By fusing hEZF to the DNA-binding domain of GAL4, we mapped a repression domain in hEZF to amino acids 181-388. We also found that amino acids 91-117 of hEZF confer an activation function on the GAL4 DNA-binding domain. |
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| Keywords |
Amino Acid Sequence; Animals; Artificial Gene Fusion; Cells, Cultured; Chromosome Mapping; Chromosomes, Human, Pair 9; Cloning, Molecular; DNA, Complementary; DNA-Binding Proteins/genetics; DNA-Binding Proteins/metabolism; Endothelium, Vascular/metabolism; Humans; Kruppel-Like Transcription Factors; Mice; Molecular Sequence Data; Sequence Homology, Amino Acid; Transcription Factors; Transcriptional Activation; Transfection; Zinc Fingers |
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Significance
Annotations
| Gene product | Qualifier | GO ID | GO term name | Evidence Code | with/from | Aspect | Notes | Status |
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