A novel complex, RUNX1-MYEF2, represses hematopoietic genes in erythroid cells.
RUNX1 is known to be an essential transcription factor for generating hematopoietic stem cells (HSC), but much less is known about its role in the downstream process of hematopoietic differentiation. RUNX1 has been shown to be part of a large transcription factor complex, together with LDB1, GATA1, TAL1, and ETO2 (N. Meier et al., Development 133:4913-4923, 2006) in erythroid cells. We used a tagging strategy to show that RUNX1 interacts with two novel protein partners, LSD1 and MYEF2, in erythroid cells. MYEF2 is bound in undifferentiated cells and is lost upon differentiation, whereas LSD1 is bound in differentiated cells. Chromatin immunoprecipitation followed by sequencing (ChIP-seq) and microarray expression analysis were used to show that RUNX1 binds approximately 9,000 target sites in erythroid cells and is primarily active in the undifferentiated state. Functional analysis shows that a subset of the target genes is suppressed by RUNX1 via the newly identified partner MYEF2. Knockdown of Myef2 expression in developing zebrafish results in a reduced number of HSC.
Department of Cell Biology and Center for Biomedical Genetics, Erasmus Medical Center, Rotterdam, Netherlands.
Boet
B
van Riel
Tibor
T
Pakozdi
Rutger
R
Brouwer
Rui
R
Monteiro
Kapil
K
Tuladhar
Vedran
V
Franke
Jan Christian
JC
Bryne
Ruud
R
Jorna
Erik-Jan
EJ
Rijkers
Wilfred
W
van Ijcken
Charlotte
C
Andrieu-Soler
Jeroen
J
Demmers
Roger
R
Patient
Eric
E
Soler
Boris
B
Lenhard
Frank
F
Grosveld
D000818:Animals; D045744:Cell Line, Tumor; D050676:Core Binding Factor Alpha 2 Subunit / Q000235:genetics / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D041905:Erythroid Cells / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D055785:Gene Knockdown Techniques; D006410:Hematopoiesis; D056466:Histone Demethylases; D051379:Mice; D060172:Morpholinos / Q000008:administration & dosage / Q000235:genetics; D046912:Multiprotein Complexes / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D010089:Oxidoreductases, N-Demethylating / Q000378:metabolism*; D011485:Protein Binding; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D015027:Zebrafish / Q000196:embryology; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism*
D050676:Core Binding Factor Alpha 2 Subunit; C576395:MYEF2 protein, zebrafish; D060172:Morpholinos; D046912:Multiprotein Complexes; C093729:Myef2 protein, mouse; D009419:Nerve Tissue Proteins; D012097:Repressor Proteins; D029961:Zebrafish Proteins; D004247:DNA; D056466:Histone Demethylases; C504076:KDM1a protein, mouse; D010089:Oxidoreductases, N-Demethylating