RRC ID 70494
Author Ryoden Y, Segawa K, Nagata S.
Title Requirement of Xk and Vps13a for the P2X7-mediated phospholipid scrambling and cell lysis in mouse T cells.
Journal Proc Natl Acad Sci U S A
Abstract A high extracellular adenosine triphosphate (ATP) concentration rapidly and reversibly exposes phosphatidylserine (PtdSer) in T cells by binding to the P2X7 receptor, which ultimately leads to necrosis. Using mouse T cell transformants expressing P2X7, we herein performed CRISPR/Cas9 screening for the molecules responsible for P2X7-mediated PtdSer exposure. In addition to Eros, which is required for the localization of P2X7 to the plasma membrane, this screening identified Xk and Vps13a as essential components for this process. Xk is present at the plasma membrane, and its paralogue, Xkr8, functions as a phospholipid scramblase. Vps13a is a lipid transporter in the cytoplasm. Blue-native polyacrylamide gel electrophoresis indicated that Xk and Vps13a interacted at the membrane. A null mutation in Xk or Vps13a blocked P2X7-mediated PtdSer exposure, the internalization of phosphatidylcholine, and cytolysis. Xk and Vps13a formed a complex in mouse splenic T cells, and Xk was crucial for ATP-induced PtdSer exposure and cytolysis in CD25+CD4+ T cells. XK and VPS13A are responsible for McLeod syndrome and chorea-acanthocytosis, both characterized by a progressive movement disorder and cognitive and behavior changes. Our results suggest that the phospholipid scrambling activity mediated by XK and VPS13A is essential for maintaining homeostasis in the immune and nerve systems.
Volume 119(7)
Published 2022-2-15
DOI 10.1073/pnas.2119286119
PII 2119286119
PMID 35140185
PMC PMC8851519
MeSH Adenosine Triphosphate Amino Acid Transport Systems, Neutral / genetics Amino Acid Transport Systems, Neutral / metabolism* Animals CRISPR-Cas Systems Cell Death Cell Line Gene Deletion Gene Expression Regulation / drug effects Genome-Wide Association Study HEK293 Cells Humans Mice Mice, Transgenic Mutation Phosphatidylserines / pharmacology Phospholipids / metabolism* Receptors, Purinergic P2X7 / genetics Receptors, Purinergic P2X7 / metabolism* T-Lymphocytes / physiology* Vesicular Transport Proteins / genetics Vesicular Transport Proteins / metabolism*
IF 9.412
DNA material pCMV-VSV-G (RDB04392)