Levels of E2A protein expression in B cell precursors are stage-dependent and inhibited by stem cell factor (c-kit ligand).
OBJECTIVE
The E2A-encoded proteins E47 and E12 are crucial to the development of pro-B and pre-B cells. The expression of E2A protein and mRNA during early B lymphopoiesis was determined and effects of stem cell factor (SCF; Steel factor; c-kit ligand) on E2A expression were evaluated.


METHODS
Ex vivo murine pro-B cells and pre-B cells were isolated and in vitro B cell precursors were derived after culture of bone marrow with rmIL-7. Levels of E2A proteins were determined by Western analysis and mRNA by RT-PCR. E2A expression in vitro was also assessed in cultures supplemented with IL-7 +/- recombinant murine SCF (rmSCF), insulin-like growth factor-1 (rhIGF-1), or Flt3-ligand (rhFlt3-L). Turnover of E2A proteins was assessed following cycloheximide treatment.


RESULTS
Ex vivo, pro-B cells had lower E47 protein levels than did pre-B cells but had comparable E2A mRNA levels. E2A protein, but not mRNA, levels were reduced in pro-B cells upon culture in vitro with IL-7 + rmSCF. This was associated with increased turnover of E2A proteins. In contrast, culture with IL-7 + IGF-1 or Flt3-L had minimal effects on E2A protein levels.


CONCLUSIONS
Pre-B cells expressed higher levels of E2A protein than did pro-B cells and this mainly resulted from posttranscriptional regulation. Exogenous SCF inhibited E2A protein, but not mRNA, expression in cultured B cell precursors, possibly by increasing E2A protein turnover. The capacity to respond to SCF may influence the levels of E2A during B-cell development.
Riley
Richard L
RL
Department of Microbiology and Immunology, University of Miami School of Medicine, PO Box 016960 (R138), Miami, FL 33101, USA. rriley@med.miami.edu
Knowles
Jennean
J
King
Anne M
AM
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002455:Cell Division / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D008565:Membrane Proteins / Q000494:pharmacology; D051379:Mice; D018345:Mice, Knockout; D019089:Stem Cell Factor / Q000494:pharmacology*; D051820:TCF Transcription Factors; D058125:Transcription Factor 7-Like 1 Protein; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000378:metabolism*
D004268:DNA-Binding Proteins; D008565:Membrane Proteins; D019089:Stem Cell Factor; D051820:TCF Transcription Factors; C544913:Tcf7l1 protein, mouse; D058125:Transcription Factor 7-Like 1 Protein; D014157:Transcription Factors; C084881:flt3 ligand protein; D007334:Insulin-Like Growth Factor I