Tyrosine phosphorylation of synapsin I by Src regulates synaptic-vesicle trafficking.
Synapsins are synaptic vesicle (SV)-associated phosphoproteins involved in the regulation of neurotransmitter release. Synapsins reversibly tether SVs to the cytoskeleton and their phosphorylation by serine/threonine kinases increases SV availability for exocytosis by impairing their association with SVs and/or actin. We recently showed that synapsin I, through SH3- or SH2-mediated interactions, activates Src and is phosphorylated by the same kinase at Tyr301. Here, we demonstrate that, in contrast to serine phosphorylation, Src-mediated tyrosine phosphorylation of synapsin I increases its binding to SVs and actin, and increases the formation of synapsin dimers, which are both potentially involved in SV clustering. Synapsin I phosphorylation by Src affected SV dynamics and was physiologically regulated in brain slices in response to depolarization. Expression of the non-phosphorylatable (Y301F) synapsin I mutant in synapsin-I-knockout neurons increased the sizes of the readily releasable and recycling pools of SVs with respect to the wild-type form, which is consistent with an increased availability of recycled SVs for exocytosis. The data provide a mechanism for the effects of Src on SV trafficking and indicate that tyrosine phosphorylation of synapsins, unlike serine phosphorylation, stimulates the reclustering of recycled SVs and their recruitment to the reserve pool.
Department of Experimental Medicine, University of Genova and Istituto Nazionale di Neuroscienze, Viale Benedetto XV 3, 161632 Genova, Italy.
Mirko
M
Messa
Sonia
S
Congia
Enrico
E
Defranchi
Flavia
F
Valtorta
Anna
A
Fassio
Franco
F
Onofri
Fabio
F
Benfenati
D000199:Actins / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000378:metabolism; D008297:Male; D008564:Membrane Potentials; D009154:Mutation; D009474:Neurons / Q000166:cytology / Q000502:physiology; D010766:Phosphorylation; D011485:Protein Binding; D055503:Protein Multimerization; D051381:Rats; D017207:Rats, Sprague-Dawley; D016704:Synapsins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013572:Synaptic Vesicles / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism*; D019061:src-Family Kinases / Q000378:metabolism*
D000199:Actins; D016704:Synapsins; D014443:Tyrosine; D019061:src-Family Kinases