An ex vivo model to study v-Myb-induced leukemogenicity.
The v-myb(AMV) oncogene transforms myelomonocytic cells in vitro and induces acute monoblastic leukemia in chickens. We analyzed the activity of the evolutionarily conserved PEST-like domain (P1 domain) for biochemical and biological activities of v-Myb in ex vivo cultures and in vivo. Deletion of the P1 domain did not affect v-Myb transcriptional activity, intracellular stability, or subcellular localization. However, it resulted in subtle yet important changes in biological activities. Although the mutant DeltaP1 v-Myb protein blocked the terminal differentiation of the monocyte/macrophage lineage as efficiently as the wild type (wt) in ex vivo cultures, it failed to induce the acute phase of monoblastic leukemia, with its fatal consequences, in vivo. Interestingly, in DeltaP1 v-myb-infected animals large numbers of monoblasts, comparable to those induced by wt v-myb, were present in the bone marrow but very few were found in the peripheral blood. The comparison of ex vivo wt- and DeltaP v-Myb bone marrow cells revealed several important features of v-Myb transformation: (i) the proliferation of transformed monoblasts is not an apparent consequence of the differentiation block with these processes being at least in part independent; (ii) the P1 domain is required for proliferation of v-Myb-mediated transformed monoblasts; (iii) the mechanism which renders transformed cells growth factor independent does not involve activation of an autocrine growth factor loop; and (iv) deletion of the P1 domain affects self-adhesion properties of v-myb-transformed monoblasts as well as their interaction with bone marrow stromal cells. These data indicate that the DeltaP1 v-myb mutant and ex vivo bone marrow cell cultures represent a valuable tool for studies on the mechanisms of leukemia formation.
Dvorakova
M
M
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Flemingovon. 2, Prague 6, 166 37, Czech Republic.
Kralova
J
J
Karafiat
V
V
Bartunek
P
P
Dvorak
M
M
D000595:Amino Acid Sequence; D000818:Animals; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D002478:Cells, Cultured; D004195:Disease Models, Animal; D020570:Genes, myb; D007938:Leukemia / Q000209:etiology / Q000235:genetics* / Q000473:pathology; D008969:Molecular Sequence Data; D009000:Monocytes / Q000473:pathology