Nitric oxide regulates endocytosis by S-nitrosylation of dynamin.
The GTPase dynamin regulates endocytic vesicle budding from the plasma membrane, but the molecular mechanisms involved remain incompletely understood. We report that dynamin, which interacts with NO synthase, is S-nitrosylated at a single cysteine residue (C607) after stimulation of the beta(2) adrenergic receptor. S-nitrosylation increases dynamin self-assembly and GTPase activity and facilitates its redistribution to the membrane. A mutant protein bearing a C607A substitution does not self-assemble properly or increase its enzymatic activity in response to NO. In NO-generating cells, expression of dynamin C607A, like the GTPase-deficient dominant-negative K44A dynamin, inhibits both beta(2) adrenergic receptor internalization and bacterial invasion. Furthermore, exogenous or endogenously produced NO enhances internalization of both beta(2) adrenergic and epidermal growth factor receptors. Thus, NO regulates endocytic vesicle budding by S-nitrosylation of dynamin. Collectively, our data suggest a general NO-dependent mechanism by which the trafficking of receptors may be regulated and raise the idea that pathogenic microbes and viruses may induce S-nitrosylation of dynamin to facilitate cellular entry.
Wang
Gaofeng
G
Department of Surgery, Howard Hughes Medical Institute, Duke University Medical Center, Durham, NC 27710, USA.
Moniri
Nader H
NH
Ozawa
Kentaro
K
Stamler
Jonathan S
JS
Daaka
Yehia
Y
D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D034281:Dynamins / Q000737:chemistry* / Q000378:metabolism; D004705:Endocytosis; D004926:Escherichia coli / Q000378:metabolism; D005434:Flow Cytometry; D005799:Genes, Dominant; D006160:Guanosine Triphosphate / Q000737:chemistry; D006801:Humans; D006868:Hydrolysis; D015151:Immunoblotting; D047468:Immunoprecipitation; D008055:Lipids / Q000737:chemistry; D009154:Mutation; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism*; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D009584:Nitrogen / Q000737:chemistry*; D011485:Protein Binding; D018343:Receptors, Adrenergic, beta-2 / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D013997:Time Factors
D008055:Lipids; D018343:Receptors, Adrenergic, beta-2; D011994:Recombinant Proteins; D009569:Nitric Oxide; D006160:Guanosine Triphosphate; D052250:Nitric Oxide Synthase Type III; D034281:Dynamins; D009584:Nitrogen; D019331:NG-Nitroarginine Methyl Ester