HOXA9 Forms Triple Complexes with PBX2 and MEIS1 in Myeloid Cells

Abstract

Aberrant activation of the HOX, MEIS, and PBX homeodomain protein families is associated with leukemias, and retrovirally driven coexpression of HOXA9 and MEIS1 is sufficient to induce myeloid leukemia in mice. Previous studies have demonstrated that HOX-9 and HOX-10 paralog proteins are unique among HOX homeodomain proteins in their capacity to form in vitro cooperative DNA binding complexes with either the PBX or MEIS protein. Furthermore, PBX and MEIS proteins have been shown to form in vivo heterodimeric DNA binding complexes with each other. We now show that in vitro DNA site selection for MEIS1 in the presence of HOXA9 and PBX yields a consensus PBX-HOXA9 site. MEIS1 enhances in vitro HOXA9-PBX protein complex formation in the absence of DNA and forms a trimeric electrophoretic mobility shift assay (EMSA) complex with these proteins on an oligonucleotide containing a PBX-HOXA9 site. Myeloid cell nuclear extracts produce EMSA complexes which appear to contain HOXA9, PBX2, and MEIS1, while immunoprecipitation of HOXA9 from these extracts results in coprecipitation of PBX2 and MEIS1. In myeloid cells, HOXA9, MEIS1, and PBX2 are all strongly expressed in the nucleus, where a portion of their signals are colocalized within nuclear speckles. However, cotransfection of HOXA9 and PBX2 with or without MEIS1 minimally influences transcription of a reporter gene containing multiple PBX-HOXA9 binding sites. Taken together, these data suggest that in myeloid leukemia cells MEIS1 forms trimeric complexes with PBX and HOXA9, which in turn can bind to consensus PBX-HOXA9 DNA targets.

Enforced Expression of NUP98-HOXA9 in Human CD34+ Cells Enhances Stem Cell Proliferation

“…In normal CD34 + cells, HOXA9 is preferentially expressed in a subfraction enriched for hematopoietic stem cells (HSC). HOXA9 has been shown to bind DNA cooperatively with either PBX1 or Meis1, two other members of the homeodomain family (12). Meis1 was originally described as a HOX cofactor that alters HOX DNA-binding specificity and affinity and increases HOX transcriptional activity (12).…”

“…HOXA9 has been shown to bind DNA cooperatively with either PBX1 or Meis1, two other members of the homeodomain family (12). Meis1 was originally described as a HOX cofactor that alters HOX DNA-binding specificity and affinity and increases HOX transcriptional activity (12). Targeted disruption of HOXA9 in mice leads to reduced numbers of progenitor cells and to a profound defect in HSC (13).…”

HoxA9 induces insulin-like growth factor-1 receptor expression in B-lineage acute lymphoblastic leukemia

“…The core homeodomain DNA-binding motif recognizes the DNA sequence (T/G)NA(T/C). 3 In mammals, there are 39 different class I Hox genes organized into four clusters (A, B, C and D), located on four separate chromosomes. The clusters may be further grouped into subclasses based on their primary sequence into 13 paralog groups within each individual cluster.…”

Genetic programs regulating HSC specification, maintenance and expansion

“…Interestingly, HSCs engineered to overexpress an activated form of STAT3 (STAT3c) behave similarly to those transduced with Hoxb4, potentially suggesting the activation of a common genetic program (Oh et al, 2003). Moreover, Hox proteins interact with Pbx which itself interacts with Meis1 forming a trimeric nuclear complex involved in target gene regulation (Swift et al, 1998;Jacobs et al, 1999;Shen et al, 1999;Liu et al, 2001). The demonstration of a genetic interaction between these transcription factors in the control of fetal liver HSC self-renewal is much awaited.…”