Multiple transacnvation domains of erythroid kruppel-like factor (eklf)

Academic Article

Abstract

  • Erythroid Kruppel-like Factor (EKLF) is an erythroid specific transcription factor that plays a critical role in γ- to β-globin gene switching during development. To identify essential domains required for EKLF transactivation function, we co-transfected a human erythroleukemia cell line (K562) with an LCR Y/LUC β/CAT reporter, and an EKLF expression vector. In this assay EKLF mediates al ,000 fold induction of β/CAT expression compared to controls. To map essential transactivation domains, progressive N-terminal, and internal deletion mutants of EKLF were constructed. All EKLF mutants were expressed at wild-type levels, localized to the nucleus, and bound DNA. When the mutant EKLF proteins were tested for β/CAT activation, a novel transactivation domain was identified. This novel domain, encompassing amino acids 140-357 is sufficient for maximal β/CAT activation. A sub-domain within this region (amino acids 140-225) is essential for its activity. Interestingly, this sub-domain is functionally redundant with the amino terminal domain (1-139). Thus, EKLF has multiple potent transactivation domains that appear to function in a redundant manner.
  • Published In

  • Blood  Journal
  • Author List

  • Pandya KR; Townes TM
  • Volume

  • 96
  • Issue

  • 11 PART I