ROCK1 but not LIMK1 or PAK2 is a key regulator of apoptotic membrane blebbing and cell disassembly

• Published in: Cell death and differentiation 2020-01, Vol.27 (1), p.102-116
• Main Authors: Tixeira, Rochelle, Phan, Thanh Kha, Caruso, Sarah, Shi, Bo, Atkin-Smith, Georgia K., Nedeva, Christina, Chow, Jenny D. Y., Puthalakath, Hamsa, Hulett, Mark D., Herold, Marco J., Poon, Ivan K. H.
• Format: Article
• Language: English
• Subjects: 13/2; 13/31; 14/19; 631/45; 631/80; Animals; Apoptosis; Apoptosis - drug effects; Biochemistry; Biomedical and Life Sciences; Cell Biology; Cell Cycle Analysis; Cell Line; Cell Membrane - ultrastructure; Cricetinae; Enzyme Inhibitors - pharmacology; Genome editing; Humans; Jurkat Cells; Life Sciences; LIM kinase; Lim Kinases - antagonists & inhibitors; Lim Kinases - physiology; Morphology; Necrosis; p21-Activated Kinases - antagonists & inhibitors; p21-Activated Kinases - physiology; rho-Associated Kinases - antagonists & inhibitors; rho-Associated Kinases - physiology; Stem Cells; THP-1 Cells
• ISSN: 1350-9047; 1476-5403
• DOI: 10.1038/s41418-019-0342-5

Many cell types are known to undergo a series of morphological changes during the progression of apoptosis, leading to their disassembly into smaller membrane-bound vesicles known as apoptotic bodies (ApoBDs). In particular, the formation of circular bulges called membrane blebs on the surface of apoptotic cells is a key morphological step required for a number of cell types to generate ApoBDs. Although apoptotic membrane blebbing is thought to be regulated by kinases including ROCK1, PAK2 and LIMK1, it is unclear whether these kinases exhibit overlapping roles in the disassembly of apoptotic cells. Utilising both pharmacological and CRISPR/Cas9 gene editing based approaches, we identified ROCK1 but not PAK2 or LIMK1 as a key non-redundant positive regulator of apoptotic membrane blebbing as well as ApoBD formation. Functionally, we have established an experimental system to either inhibit or enhance ApoBD formation and demonstrated the importance of apoptotic cell disassembly in the efficient uptake of apoptotic materials by various phagocytes. Unexpectedly, we also noted that ROCK1 could play a role in regulating the onset of secondary necrosis. Together, these data shed light on both the mechanism and function of cell disassembly during apoptosis.