Phosphatidylinositol (4,5)-bisphosphate-dependent activation of dynamins I and II lacking the proline/arginine-rich domains.
Dynamins comprise a family of GTPases that participate in the early stages of endocytosis. The GTPase activity of neuronal specific dynamin I is stimulated by microtubules, negatively charged phospholipid vesicles, and Src homology 3-containing proteins, including Grb2. These activators were previously shown to bind to a proline/arginine-rich domain (PRD) in the carboxyl-terminal region of the enzyme. Dynamin II, which is ubiquitously expressed, had not been purified or characterized previously. In this study, the enzymatic properties of rat dynamin II and of D746, a dynamin II truncation mutant lacking the PRD, have been characterized. Dynamin II has a higher basal activity than dynamin I, but the two types of dynamin are stimulated similarly by microtubules, Grb2, and phospholipids. D746 is not activated by microtubules or Grb2, highlighting the significance of the PRD for these interactions, but it is activated by phospholipid vesicles containing phosphatidylserine or phosphatidylinositol-4,5- bisphosphate. Moreover, in contrast to previous reports, the PRD appears not to be required for phospholipid-stimulated self-assembly of dynamin, which is a key element in the regulation of its activity. Similar results were obtained with bovine brain dynamin I that had been subjected to limited proteolytic digestion to remove the PRD. Our data highlight the potential involvement of dynamin pleckstrin homology domains in the regulation of GTPase activity by phospholipids.
Lin
H C
HC
Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas 75235, USA.
Barylko
B
B
Achiriloaie
M
M
Albanesi
J P
JP
D000818:Animals; D001120:Arginine / Q000378:metabolism*; D001665:Binding Sites; D001923:Brain Chemistry; D002417:Cattle; D034284:Dynamin I; D034281:Dynamins; D004789:Enzyme Activation; D020558:GTP Phosphohydrolases / Q000737:chemistry / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D009994:Osmolar Concentration; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism*; D011392:Proline / Q000378:metabolism*; D051381:Rats
D019269:Phosphatidylinositol 4,5-Diphosphate; D001120:Arginine; D011392:Proline; D034284:Dynamin I; D020558:GTP Phosphohydrolases; D034281:Dynamins