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 Previous Article

Molecular and Cellular Biology, April 2007, p. 2777-2790, Vol. 27, No. 7
0270-7306/07/$08.00+0     doi:10.1128/MCB.01658-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Erythroid Krüppel-Like Factor Directly Activates the Basic Krüppel-Like Factor Gene in Erythroid Cells{triangledown}

Alister P. W. Funnell,1 Christopher A. Maloney,1,{dagger} Lucinda J. Thompson,1,{ddagger} Janelle Keys,2 Michael Tallack,2 Andrew C. Perkins,2 and Merlin Crossley1*

School of Molecular and Microbial Biosciences, G08, University of Sydney, Sydney, New South Wales 2006, Australia,1 Institute for Molecular Bioscience, University of Queensland, Brisbane, Queensland, Australia2

Received 6 September 2006/ Returned for modification 16 October 2006/ Accepted 12 January 2007

The Sp/Krüppel-like factor (Sp/Klf) family is comprised of around 25 zinc finger transcription factors that recognize CACCC boxes and GC-rich elements. We have investigated basic Krüppel-like factor (Bklf/Klf3) and show that in erythroid tissues its expression is highly dependent on another family member, erythroid Krüppel-like factor (Eklf/Klf1). We observe that Bklf mRNA is significantly reduced in erythroid tissues from Eklf-null murine embryos. We find that Bklf is driven primarily by two promoters, a ubiquitously active GC-rich upstream promoter, 1a, and an erythroid downstream promoter, 1b. Transcripts from the two promoters encode identical proteins. Interestingly, both the ubiquitous and the erythroid promoter are dependent on Eklf in erythroid cells. Eklf also activates both promoters in transient assays. Experiments utilizing an inducible form of Eklf demonstrate activation of the endogenous Bklf gene in the presence of an inhibitor of protein synthesis. The kinetics of activation are also consistent with Bklf being a direct Eklf target. Chromatin immunoprecipitation assays confirm that Eklf associates with both Bklf promoters. Eklf is typically an activator of transcription, whereas Bklf is noted as a repressor. Our results support the hypothesis that feedback cross-regulation occurs within the Sp/Klf family in vivo.

* Corresponding author. Mailing address: Merlin Crossley School of Molecular and Microbial Biosciences, G08, University of Sydney, Sydney, NSW 2006, Australia. Phone: 61-2-9351 2233. Fax: 61-2-9351 4726. E-mail: m.crossley{at}mmb.usyd.edu.au.

{triangledown} Published ahead of print on 5 February 2007.

{dagger} Present address: Division of Obesity and Metabolic Health, Rowett Research Institute, Aberdeen AB21 9SB, Scotland.

{ddagger} Present address: Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305-5124.

Molecular and Cellular Biology, April 2007, p. 2777-2790, Vol. 27, No. 7
0270-7306/07/$08.00+0     doi:10.1128/MCB.01658-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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