The amino terminal and E2F interaction domains are critical for C/EBPα-mediated induction of granulopoietic development of hematopoietic cells

• Published in: Blood 2003-11, Vol.102 (9), p.3163-3171
• Main Authors: D’Alo’, Francesco, Johansen, Lisa M., Nelson, Erik A., Radomska, Hanna S., Evans, Erica K., Zhang, Pu, Nerlov, Claus, Tenen, Daniel G.
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Binding Sites; Biological and medical sciences; CCAAT-Enhancer-Binding Protein-alpha - chemistry; CCAAT-Enhancer-Binding Protein-alpha - genetics; CCAAT-Enhancer-Binding Protein-alpha - physiology; Cell Cycle Proteins; Cell differentiation, maturation, development, hematopoiesis; Cell physiology; DNA-Binding Proteins; E2F Transcription Factors; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation; Granulocytes - cytology; Hematopoiesis; Hematopoietic Stem Cells - cytology; Humans; K562 Cells; Molecular and cellular biology; Mutation; Protein Structure, Tertiary - physiology; Transcription Factors - genetics; Transcription Factors - metabolism; Transcription Factors - physiology
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood-2003-02-0479

The transcription factor C/EBPα (CCAAT/enhancer binding protein α) is critical for granulopoiesis. Gene disruption in mice blocks early granulocyte differentiation and disruption of C/EBPα function has been implicated in human acute myeloid leukemia (AML), but no systematic structure-function analysis has been undertaken to identify the mechanisms involved in C/EBPα-mediated granulocyte differentiation. Here we demonstrate that loss of either of 2 key regions results in disruption of C/EBPα granulocytic development: the amino terminus and specific residues residing on the non-DNA binding face of the basic region. Mutation of either results in loss of C/EBPα inhibition of E2F and down-regulation of c-Myc, but only mutation of the basic region results in loss of physical interaction with E2F. In contrast, while the amino terminal mutant retains the ability to interact with E2F, this mutant fails to bind a C/EBPα site efficiently, fails to activate C/EBPα target genes, and is also defective in inhibition of E2F activity. These results further emphasize the importance of inhibition of proliferative pathways in granulopoiesis and demonstrate that several regions of the C/EBPα protein are involved in this mechanism.