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Vol. 273, Issue 2, 1026-1031, January 9, 1998

Human EZF, a Krüppel-like Zinc Finger Protein, Is Expressed in Vascular Endothelial Cells and Contains Transcriptional Activation and Repression Domains

Shaw-Fang Yet, Megan M. McA'Nulty, Sara C. Folta, Hsueh-Wei Yen, Masao Yoshizumi, Chung-Ming Hsieh, Matthew D. Layne, Michael T. ChinDagger §, Hong Wang, Mark A. PerrellaDagger , Mukesh K. JainDagger §, and Mu-En LeeDagger §

From the Cardiovascular Biology Laboratory, Harvard School of Public Health, Dagger  Department of Medicine, Harvard Medical School,  Pulmonary and § Cardiovascular Divisions, Brigham and Women's Hospital, Boston, Massachusetts 02115

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.

Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.


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