Figure 8.

Model of origin licensing dynamics in intestinal epithelial cells. In a normal cell cycle, the Mcm cells are expressed ubiquitously in all stages. The licensing of DNA with MCM2–7 occurs in late M and throughout G1, when a cell receives a stimulus to commit to the cell cycle. As DNA is replicated during the S phase, MCM2–7 are displaced from DNA and are prevented from relicensing in G2. During terminal differentiation, MCM2–7 are not actively transcribed, and the proteins are gradually lost in postmitotic cells. However, after the final mitotic division, cells make a binary decision never to license their DNA, even though the protein is still present. Mcm proteins then degrade slowly, where cells enter a terminally differentiated state (deep G0). Alternatively, cells can exit mitosis, not relicense their DNA but maintain proliferative markers, and disengage from the cell cycle for some time (unlicensed G1). Two major classes of intestinal stem cells exist: “active” stem cells, engaged with the cell cycle, and reserve, quiescent LRCs. LRCs are in a state of “deep” quiescence and do not contain MCM2–7 because they have disengaged from the cell cycle for some time. In this study, we show that most active Lgr5+ stem cells reside in an unlicensed state but contain MCM2–7 proteins. These cells reside in an unlicensed G1 phase until they make a proliferative-fate decision, enter the cell cycle, and license. This provides an explanation for the elongated cell cycle of intestinal stem cells; they reside in a partial resting state in which they may be able to respond to niche cues to divide. This, therefore, may constitute a unique mechanism to control stem cell numbers.

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