A global role for EKLF in definitive and primitive erythropoiesis

Perkins, Andrew C., Coghill, Elise, Maguire, Tina, Hartmann, Belinda, McDowall, Alasdair, Weiss, Mitchell, Grimmond, Sean, Keys, Janelle and Hodge, Denise (2005). A global role for EKLF in definitive and primitive erythropoiesis. In: Blood. ASH Annual Meeting Abstracts. 47th American Society of Hematology Annual Meeting, New Orleans, USA, (496A-496A). 3-6 December 2005.


Author Perkins, Andrew C.
Coghill, Elise
Maguire, Tina
Hartmann, Belinda
McDowall, Alasdair
Weiss, Mitchell
Grimmond, Sean
Keys, Janelle
Hodge, Denise
Title of paper A global role for EKLF in definitive and primitive erythropoiesis
Conference name 47th American Society of Hematology Annual Meeting
Conference location New Orleans, USA
Conference dates 3-6 December 2005
Proceedings title Blood. ASH Annual Meeting Abstracts   Check publisher's open access policy
Place of Publication Washington, D.C., USA
Publisher American Society of Hematology
Publication Year 2005
Sub-type Published abstract
ISSN 0006-4971
1528-0020
Volume 106
Issue 11
Start page 496A
End page 496A
Language eng
Abstract/Summary Email Alerts Blood (ASH Annual Meeting Abstracts) 2005 106: Abstract 1745 © 2005 American Society of Hematology This Article Services Right arrow Email this article to a friend Right arrow Download to citation manager Citing Articles Right arrow Citing Articles via Google Scholar Google Scholar Right arrow Articles by Perkins, A. C. Right arrow Articles by Hodge, D. Right arrow Search for Related Content PubMed Right arrow Articles by Perkins, A. C. Right arrow Articles by Hodge, D. Social Bookmarking Add to CiteULike Add to Connotea Add to Del.icio.us Add to Digg Add to Reddit Add to Technorati What's this? Poster Sessions A Global Role for EKLF in Definitive and Primitive Erythropoiesis. Andrew C. Perkins, MBBS, PhD1,*, Elise Coghill, PhD1,*, Tina Maguire, PhD1,*, Belinda Hartmann, BSc1,*, Alasdair McDowall, PhD1,*, Mitchell Weiss, MD, PhD2, Sean Grimmond, PhD1,*, Janelle Keys, PhD1,* and Denise Hodge, PhD1,* 1 Insitute for Molecular Bioscience, University of Queensland, St Lucia, Queensland, Australia and 2 The Children’s Hospital of Philadelphia, University of Pennsylvania, Philadelphia, PA, USA. Abstract Erythroid Kruppel-like factor (EKLF or Klf1) is an erythroid specific C2H2 zinc-finger transcription factor which is essential for definitive erythropoiesis and ß-globin gene expression. The absence of EKLF results in fatal anaemia but correction of globin chain imbalance does result in rescue, suggesting the existence of additional EKLF target genes. The aim of this study was to search for such genes by expression profiling. We performed profiling on fetal livers from wild-type versus EKLF null litter mates, and also EKLF null erythroid cell lines containing an inducible EKLF-ERTM fusion construct. Hybridisations were performed on microarray slides printed with a 23K oligo library from Compugen. Target gene validation was performed by real-time RT-PCR, chromatin immuno-precipitation (ChIP) and promoter-reporter assays. A large number of genes were down regulated in the absence of EKLF but few were up regulated, suggesting EKLF acts primarily as a transcriptional activator in vivo. One hundred genes were EKLF dependent in both systems. These include heme synthesis enzymes, red cell surface proteins including Rh and the transferrin receptor, and erythroid transcription factors. Two interesting highly EKLF-dependent genes are {alpha}-haemoglobin stabilising protein (AHSP), a key chaperone for free a-globin chains, and dematin (band 4.9) which links the cytoskeleton to the red cell membrane. A search for EKLF binding sites within the dematin and AHSP genes demonstrated a number of phylogenetically conserved CACC sites, and ChIP demonstrated in vivo EKLF occupancy at some but not all of these. Promoter-reporter assays showed EKLF directly activates dematin gene transcription through two promoters containing these sites. Lastly, investigation of EKLF target genes in the yolk sac lead to the discovery of unexpected defects in the embryonic red cell membrane and cytoskeleton. In conclusion, EKLF regulates global erythroid gene expression which is critical for development of primitive as well as definitive red cells.
Subjects C1
270201 Gene Expression
730103 Blood disorders
Keyword Hematology
Kruppel-like Factor
Transcription Factor Gata-1
5-aminolevulinate Synthase Gene
Porphobilinogen Deaminase Gene
Box-binding-protein
Beta-thalassemia
Gamma-globin
Sideroblastic Anemia
Histone Acetylation
Regulatory Elements
Q-Index Code EX
Additional Notes Abstract number: 1745

 
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Created: Tue, 02 Mar 2010, 13:33:15 EST by Laura McTaggart on behalf of Institute for Molecular Bioscience