Transient Primary Cilia Mediate Robust Hedgehog Pathway-Dependent Cell Cycle Control.

The regulation of proliferation is a primary function of Hedgehog (Hh) signaling in development. Hh signal transduction requires the primary cilium for several steps in the pathway [1-5]. Many cells only build a primary cilium upon cell cycle exit, in G0. In those proliferating cells that do make a cilium, it is a transient organelle, being assembled in G1 and disassembled sometime prior to mitosis [6-9]. Thus, the requirement for primary cilia presents a conundrum: how are proliferative signals conveyed through an organelle that is present for only part of the cell cycle? Here, we investigate this question in a mouse medulloblastoma cell line, SMB55, that requires cilium-mediated Hh pathway activity for proliferation [10]. We show that SMB55 cells, and the primary cerebellar granule neuron precursors (GNPs) from which they derive, are often ciliated beyond G1 into S phase, and the presence of the cilium in SMB55 cells determines the periods of Hh pathway activity. Using live imaging over multiple cell cycles, we demonstrate that Hh pathway activity in either G1-S of the previous cell cycle or G1 of the cell cycle in which the decision is made is sufficient for cell cycle entry. We also show that cyclin D1 contributes to the persistent effects of pathway activity over multiple cell cycles. Together, our results reveal that, even though the signaling organelle itself is transient, Hh pathway control of proliferation is remarkably robust. Further, primary cilium transience may have implications for other Hh-mediated events in development.