The translocation of focal adhesion kinase in brain synaptosomes is regulated by phosphorylation and actin assembly.
Focal adhesion kinase (FAK) and the related proline-rich tyrosine kinase 2 (PYK2) are non-receptor protein tyrosine kinases that transduce extracellular signals through the activation of Src family kinases and are highly enriched in neurones. To further elucidate the regulation of FAK and PYK2 in nervous tissue, we investigated their distribution in brain subcellular fractions and analysed their translocation between membrane and cytosolic compartments. We have found that FAK and PYK2 are present in a small membrane-associated pool and a larger cytosolic pool in various neuronal compartments including nerve terminals. In intact nerve terminals, inhibition of Src kinases inhibited the membrane association of FAK, but not of PYK2, whereas tyrosine phosphatase inhibition sharply increased the membrane association of both FAK and PYK2. Disruption of the actin cytoskeleton was followed by a decrease in the membrane-associated pool of FAK, but not of PYK2. For both kinases, a significant correlation was found between autophosphorylation and membrane association. The data indicate that FAK and PYK2 are present in nerve terminals and that the membrane association of FAK is regulated by both phosphorylation and actin assembly, whereas that of PKY2 is primarily dependent on its phosphorylation state.
Bongiorno-Borbone
Lucilla
L
Department of Experimental Medicine, Section of Human Physiology, University of Genova, Viale Benedetto, Genova, Italy.
Onofri
Franco
F
Giovedì
Silvia
S
Ferrari
Rosaria
R
Girault
Jean-Antoine
JA
Benfenati
Fabio
F
D000199:Actins / Q000502:physiology*; D000818:Animals; D001692:Biological Transport / Q000502:physiology; D001921:Brain / Q000201:enzymology*; D003599:Cytoskeleton / Q000502:physiology; D051417:Focal Adhesion Kinase 1; D051418:Focal Adhesion Kinase 2; D051416:Focal Adhesion Protein-Tyrosine Kinases; D007425:Intracellular Membranes / Q000378:metabolism; D010766:Phosphorylation; D016548:Prosencephalon / Q000201:enzymology; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012995:Solubility; D013347:Subcellular Fractions / Q000201:enzymology; D013574:Synaptosomes / Q000201:enzymology*; D014443:Tyrosine / Q000378:metabolism; D018909:src Homology Domains / Q000502:physiology
D000199:Actins; D014443:Tyrosine; D011505:Protein-Tyrosine Kinases; D051417:Focal Adhesion Kinase 1; D051418:Focal Adhesion Kinase 2; D051416:Focal Adhesion Protein-Tyrosine Kinases; C496052:Ptk2 protein, rat; C496054:Ptk2b protein, rat