10779/crick.12652940.v1
Ianina L Conte
Ianina L
Conte
Nicola Hellen
Nicola
Hellen
Ruben Bierings
Ruben
Bierings
Gregory I Mashanov
Gregory I
Mashanov
Jean-Baptiste Manneville
Jean-Baptiste
Manneville
Nikolai I Kiskin
Nikolai I
Kiskin
Matthew J Hannah
Matthew J
Hannah
Justin E Molloy
Justin E
Molloy
Tom Carter
Tom
Carter
Interaction between MyRIP and the actin cytoskeleton regulates Weibel-Palade body trafficking and exocytosis
The Francis Crick Institute
2020
Actin
Exocytosis
MyRIP
Myosin Va
Weibel–Palade body
Actin Cytoskeleton
Actins
Calcium
Cell Line
Cell Movement
Green Fluorescent Proteins
Human Umbilical Vein Endothelial Cells
Humans
Myosin Heavy Chains
Myosin Type V
Protein Binding
Protein Transport
Vesicular Transport Proteins
Weibel-Palade Bodies
Molloy FC001119
Schaefer FC001153
06 Biological Sciences
11 Medical and Health Sciences
Developmental Biology
2020-07-15 11:15:21
Journal contribution
https://crick.figshare.com/articles/journal_contribution/Interaction_between_MyRIP_and_the_actin_cytoskeleton_regulates_Weibel-Palade_body_trafficking_and_exocytosis/12652940
Weibel-Palade body (WPB)-actin interactions are essential for the trafficking and secretion of von Willebrand factor; however, the molecular basis for this interaction remains poorly defined. Myosin Va (MyoVa or MYO5A) is recruited to WPBs by a Rab27A-MyRIP complex and is thought to be the prime mediator of actin binding, but direct MyRIP-actin interactions can also occur. To evaluate the specific contribution of MyRIP-actin and MyRIP-MyoVa binding in WPB trafficking and Ca(2+)-driven exocytosis, we used EGFP-MyRIP point mutants with disrupted MyoVa and/or actin binding and high-speed live-cell fluorescence microscopy. We now show that the ability of MyRIP to restrict WPB movement depends upon its actin-binding rather than its MyoVa-binding properties. We also show that, although the role of MyRIP in Ca(2+)-driven exocytosis requires both MyoVa- and actin-binding potential, it is the latter that plays a dominant role. In view of these results and together with the analysis of actin disruption or stabilisation experiments, we propose that the role of MyRIP in regulating WPB trafficking and exocytosis is mediated largely through its interaction with actin rather than with MyoVa.