REGULATION OF G(1)/S TRANSITION BY CYCLIN-D2 AND CYCLIN-D3 IN HEMATOPOIETIC-CELLS

Identification of the genes that control passage through the G1 phase of the cell cycle in mammalian cells is of particular interest because virtually all external events that regulate proliferation act primarily or exclusively during G1. Cyclins are likely to play a key role in controlling cell cycle progression, although their role during G1 in higher eukaryotic cells is unclear. In the hematopoietic cell line 32Dcl3, both cyclins D2 and D3 were expressed in proliferating cells, while cyclin D1 was undetectable. Expression of D2, and to a lesser extent D3, was interleukin 3 (IL-3) dependent and declined rapidly in the absence of this growth factor. To investigate the potential role of D cyclins in regulating cell growth, cell lines overexpressing either D2 or D3 were generated by transfection. Constitutive overexpression of either D2 or D3 did not affect cell viability, rate of cell proliferation, or dependence on IL-3 for growth. However, the distribution of cells through the cell cycle was dramatically altered, with both cyclins causing an increase in the fraction of cells in S phase, apparently related to a shortening of G1. Also, when deprived of IL-3, D3-overexpressing cells failed to arrest in G1, and apoptotic cell death in the absence of IL-3 was delayed. These results suggest a role for cyclins D2 and D3 in controlling passage of hematopoietic cells through G1 in the presence of growth factors and in effecting G1 arrest in the absence of growth factors.