Genesis, a Winged Helix Transcriptional Repressor with Expression Restricted to Embryonic Stem Cells

doi:10.1074/jbc.271.38.23126

Genesis, a Winged Helix Transcriptional Repressor with Expression Restricted to Embryonic Stem Cells*

From the ^‡ Division of Hematology/Oncology and the Walther Oncology Center, IB 442, Indiana University Medical Center, Indianapolis, Indiana 46202-5121, the
^§ Department of Biochemistry, University of Illinois College of Medicine, Chicago, Illinois 60612-7334,
^∥ Krannert Institute of Cardiology, Indiana University Medical Center, Indianapolis, Indiana 46202,
^‴ Mammalian Genetics Laboratory, ABL-Basic Research Program, NCI, National Institutes of Health, Frederick Cancer Research and Development Center, Frederick, Maryland 21702, and the
¹ Department of Microbiology/Immunology, MS 252, Indiana University Medical Center, Indianapolis, Indiana 46202-5120

^∥ A Scholar of the Leukemia Society of America, supported by USPHS Grant HL 48915. To whom correspondence should be addressed:
Div. of Hematology/Oncology and the Walther Oncology Center, IB 442, Indiana University Medical Center, 975 W. Walnut St., Indianapolis, IN 46202-5121.
Tel.: 317-274-3589. Fax: 317-274-0396. E-mail: robert_ hromas{at}iucc.iupui.edu

Abstract

A novel member of the winged helix (formerly HNF-3/Forkhead) transcriptional regulatory family, termed Genesis, was isolated and characterized. Putative translation of the complete cDNA revealed the winged helix DNA binding domain to be centrally located within the protein, with regions on either side that contain known transcriptional regulatory motifs. Extensive Northern analysis of Genesis found that the message was exclusively expressed in embryonic stem cells or their malignant equivalent, embryonal carcinoma cells. The Genesis transcript was down-regulated when these cells were stimulated to differentiate. DNA sequences that Genesis protein would interact with were characterized and were found to contain a consensus similar to that found in an embryonic stem cell enhancer sequence. Co-transfection experiments revealed that Genesis is a transcriptional repressor. Genesis mapped to mouse chromosome 4 in a region syntenic with human chromosome 1p31, a site of nonrandom abnormalities in germ cell neoplasia, neuroblastoma, and acute lymphoblastic leukemia. Genesis is a candidate for regulating the phenotype of normal or malignant embryonic stem cells.

Footnotes

↵^¶ An Established Investigator of the American Heart Association/Bristol Myers Squibb, supported by United States Public Health Service (USPHS) Grant GM43241 from the National Institutes of Health.
^‴ A Fellow of the Leukemia Society of America.
↵* Portions of this work were supported by NCI, National Institutes of Health, under contract with ABL. 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.
↵¹ The abbreviations used are:

HNF

hepatocyte nuclear factor

HFH

HNF-3/Forkhead

CAT

chloramphenicol acetyltransferase

FKH

Forkhead

kb

kilobase pair(s).
- Received April 26, 1996.
- Revision received June 27, 1996.