Identification of a series of differentiation-associated gene sequences from GM-CSF stimulated bone marrow.
The production of terminally differentiated granulocytes and monocytes occurs by a program of orderly and sequential gene expression. This genetic program can be studied in vitro utilizing granulocyte-monocyte colony stimulating factor (GM-CSF) to stimulate proliferation and myeloid cell maturation. We have identified a series of genes expressed during early myeloid differentiation by differential screening of a cDNA library prepared from GM-CSF stimulated murine progenitor cells. From 72 potential early myeloid specific clones, three (B9, C9, C15) were characterized further. The time course of RNA accumulation during GM-CSF stimulated maturation demonstrated a unique pattern for each clone. B9 was expressed predominantly on day 3, and followed a pattern of accumulation similar to that for myeloperoxidase. (Jaffe et al. (1988), Oncogene, 2, 167-174). C9 expression increased gradually to a maximum level on day 3 and then plateaued; it appeared to be equally expressed in granulocytes and monocytes. C15 was expressed at a consistently high level in unstimulated cells and at 1-3 days post GM-CSF stimulation. It then decreased to undetectable levels. Hybridization of these clones to a panel of murine tissue RNAs demonstrated restricted expression of B9 to bone marrow, while C9 was present in most murine tissues, including thymus. C15 expression was relatively restricted to ovary/uterus, liver and adrenal, in addition to bone marrow. Partial DNA sequence analysis suggested that B9 was a novel sequence not previously identified. C9 was identified as thymosin beta-4, and C15 showed extensive homology to lactotransferrin. Thus, screening a bone marrow cDNA library by differential hybridization has successfully yielded a series of DNA sequences regulated during murine myelopoiesis. These include novel sequences (B9), genes previously known to be regulated during myelopoiesis ( C15), as well as sequences not recognized previously as being associated with myeloid differentiation (C9).[1]References
- Identification of a series of differentiation-associated gene sequences from GM-CSF stimulated bone marrow. Moscinski, L.C., Prystowsky, M.B. Oncogene (1990) [Pubmed]
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