An unmet actin requirement explains the mitotic inhibition of clathrin-mediated endocytosis

Abstract

Clathrin-mediated endocytosis (CME) is the major internalisation route for many different receptor types in mammalian cells. CME is shut down during early mitosis, but the mechanism of this inhibition is unclear. In this study, we show that the mitotic shutdown is due to an unmet requirement for actin in CME. In mitotic cells, membrane tension is increased and this invokes a requirement for the actin cytoskeleton to assist the CME machinery to overcome the increased load. However, the actin cytoskeleton is engaged in the formation of a rigid cortex in mitotic cells and is therefore unavailable for deployment. We demonstrate that CME can be ‘restarted’ in mitotic cells despite high membrane tension, by allowing actin to engage in endocytosis. Mitotic phosphorylation of endocytic proteins is maintained in mitotic cells with restored CME, indicating that direct phosphorylation of the CME machinery does not account for shutdown.The plasma membrane that surrounds a cell acts as a protective barrier that regulates what can enter or exit the cell. However, large molecules and other ‘cargo’ can get into a cell in a variety of ways. One of these routes—known as clathrin-mediated endocytosis—involves a receptor on the outside of the membrane grabbing hold of the cargo while a protein called clathrin forms a ‘pit’ beneath the receptor. This pit becomes deeper and deeper until the cargo is completely surrounded by clathrin-lined membrane and is brought inside the cell.This process has been studied over the past 50 years, and it is known that clathrin-mediated endocytosis is turned off when a cell begins to divide to produce new cells, and then turned back on when cell division has come to an end. However, there are competing theories as to exactly why this process stops when cell division starts.Now, Kaur et al. have investigated these theories by looking at the role that another protein, called actin, plays in turning off clathrin-mediated endocytosis. Actin is a molecule that forms a sort of scaffolding within the cell (called the cytoskeleton), and it also guides the movement of molecules and larger structures within the cell. Further, when the cell membrane is being stretched, the actin cytoskeleton can assist the clathrin-mediated endocytosis machinery to pull cargo into the cell.So why doesn’t actin help with endocytosis during cell division? The answer, Kaur et al. suggest, is that all the actin in the cell is needed by the cytoskeleton during cell division, so there is no actin available to perform other tasks such as clathrin-mediated endocytosis. Further experiments demonstrated that this form of endocytosis can be ‘restarted’ in dividing cells by treating the cells in a way that frees up some additional actin. The work of Kaur et al. also ruled out the theory that chemical changes to the endocytosis machinery disabled it during cell division. These findings have implications for the delivery of drugs, via endocytosis, to the rapidly dividing cells that are involved in diseases such as cancer.