Cloning the cDNA for a New Human Zinc Finger Protein Defines a Group of Closely Related Krüppel-like Transcription Factors

doi:10.1074/jbc.273.43.28229

Cloning the cDNA for a New Human Zinc Finger Protein Defines a Group of Closely Related Krüppel-like Transcription Factors*

From the ^‡Genetics Division, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-Ku, Tokyo 104-0045, Japan and the ^§Brookdale Center for Developmental and Molecular Biology, Mount Sinai School of Medicine, New York, New York 10029

Abstract

We have identified a novel zinc finger protein that has been named ubiquitous Krüppel-like factor (UKLF) based on structural considerations and the pattern of gene expression. UKLF was isolated by the polymerase chain reaction approach using degenerate oligonucleotides corresponding to the DNA-binding domain of erythroid Krüppel-like factor (EKLF) and cDNA prepared from human vascular endothelial cells. The carboxyl-terminal portion of UKLF contains three zinc fingers of the Cys₂-His₂ type and binds in vitro to the CACCC motif of the β-globin promoter and to the Sp1 recognition sequence. The amino-terminal portion of UKLF consists of a hydrophobic region rich in serines and a negatively charged segment with several glutamic acid residues. The first 47 amino acids of the acidic region are nearly identical to the amino-terminal portion of another Krüppel-like factor, the so-called core promoter-binding protein (CPBP) or Zf9. Like CPBP/Zf9, UKLF can function as a transcription activator in co-transfection assays. However, this activity is lost when the highly conserved amino-terminal segment is deleted. These findings indicate that UKLF and CPBP/Zf9 represent a distinct subgroup of closely related Krüppel-like activators of transcription. Mapping of the UKLF gene to chromosome 2 suggested that UKLF and CPBP/Zf9 translocated to different chromosomes following duplication from an ancestral gene.

Footnotes

↵* This work was supported by a grant-in-aid for the 2nd Term Comprehensive 10-Year Strategy for Cancer Control and Health Sciences Research Grants from the Ministry of Health and Welfare of Japan, by the Mitsui Life Social Welfare Foundation, and by Grants AR-38648 and AR-42841 from the National Institutes of Health. This is article 268 from the Brookdale Center for Developmental and Molecular Biology.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) AB015132.
↵¶ To whom correspondence should be addressed: Brookdale Center for Developmental and Molecular Biology, Mount Sinai School of Medicine, One Gustave L. Levy Pl., Box 1126, New York, NY 10029. Tel.: 212-241-7984; E-mail: ramirf01{at}doc.mssm.edu.
↵2 Laub et al., manuscript in preparation.
Abbreviations:

BKLF

basic Krüppel-like factor

BTEB2

basic transcription element binding protein 2

CPBP

core promoter binding protein

CAT

chloramphenicol acetyltransferase

EMSA

electrophoretic mobility shift assay

GKLF

gut Krüppel-like factor

EKLF

erythroid Krüppel-like factor

FISH

fluorescence in situ hybridization

GFP

green fluorescent protein

LKLF

lung Krüppel-like factor

LUC

luciferase

ORF

open reading frame

NLS

nuclear localization signal

RT

reverse transcriptase

PCR

polymerase chain reaction

RACE

rapid amplification of cDNA ends

TK

thymidine kinase

UKLF

ubiquitous Krüppel-like factor

HUVEC

human umbilical vein endothelial cell

PSG

pregnancy-specific glycoprotein

DAPI

4′,6-diamidino-2-phenylindole.
- Received June 26, 1998.
- Revision received August 4, 1998.
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