Nucleic Acids Research, Vol 27, Issue 24 4807-4815, Copyright © 1999 by Oxford University Press
H Shi, Z Zhang, X Wang, S Liu and CT Teng
The mouse lactoferrin gene promoter includes a CAAT/GT box,
GGGCAATAGGGTGGGGCCAGCCC, which functions as the epidermal growth factor
response element (EGFRE) in human endometrial carcinoma RL95-2 cells
(RL95). A positive clone, EGFREB, of 2575 bp length, was isolated from an
expression library of RL95 cells with a multimer of the EGFRE sequence. In
this work, we have identified that EGFREB encodes the C- terminus of
Kruppel-like factor 5 (KLF5). This mRNA is most abundant in human colon and
small intestine. A full-length cDNA clone was isolated from a human colon
library using EGFREB as the hybridization probe. The full-length cDNA
consists of 3336 bp with a 302 bp 5'-UTR, a 1663 bp 3'- UTR, and a 1371 bp
sequence coding for a 457 amino acid polypeptide. Based on its tissue
distribution and sequence homology to the mouse IKLF, we renamed this
protein IKLF. DNase I footprinting and electrophoresis mobility shift assay
confirmed the binding of IKLF to the EGFRE. The human IKLF gene spans
>20 kb in length and is organized into four exons, whose intron/exon
junctions follow the GT/AG rule. The three zinc fingers are encoded by
three exons. Nuclear localization of IKLF was demonstrated by green
fluorescence protein (GFP)-tagged IKLF in transfection experiments and
western analysis. Overexpression of IKLF in RL95 cells represses the
activity of reporter constructs containing the CAAT/GT box of the mouse
lactoferrin gene. These findings imply that IKLF is a nuclear transcription
factor that binds to the CAAT/GT box, and functions as a modulator of the
mouse lacto- ferrin gene promoter activity.
ARTICLES
Isolation and characterization of a gene encoding human Kruppel-like factor 5 (IKLF): binding to the CAAT/GT box of the mouse lactoferrin gene promoter
Gene Regulation Group, Laboratory of Reproductive and Development Toxicology, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC 27709, USA.
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