Microtubule/MAP-affinity regulating kinase (MARK) is activated by phenylarsine oxide in situ and phosphorylates tau within its microtubule-binding domain.
Tau is a microtubule-associated protein (MAP) that is functionally modulated by phosphorylation and that is hyperphosphorylated in several neurodegenerative diseases. Because phosphorylation regulates both normal and pathological tau functioning, it is of interest to identify the signaling pathways and enzymes capable of modulating tau phosphorylation in vivo. Previously, it was demonstrated that in SH-SY5Y human neuroblastoma cells and rat primary cortical cultures tau is phosphorylated at Ser262/356, within its microtubule-binding domain, by a staurosporine-sensitive protein kinase in response to the vicinal thiol-directed agent phenylarsine oxide. The current study demonstrates the presence of a 100-kDa protein kinase activity in SH-SY5Y cells that associates with microtubules, phosphorylates tau at Ser262/356, is activated by phenylarsine oxide, and is inhibited by the protein kinase inhibitor staurosporine. Isolation of individual protein bands from a polyacrylamide gel revealed two closely spaced proteins containing Ser262/356-directed protein kinase activity. Mass spectrometry analysis indicated that these protein bands correspond to the 100-kDa microtubule/MAP-affinity regulating kinase (MARK), which has been shown previously to phosphorylate tau within its microtubule-binding domain. Immunoblot analysis of the protein kinase bands confirmed this finding, providing the first demonstration that activation of endogenous MARK results in increased tau phosphorylation within its microtubule-binding domain in situ.
Jenkins
S M
SM
Department of Psychiatry, University of Alabama at Birmingham, 35294-0017, USA. gvwj@uab.edu
Johnson
G V
GV
D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D001152:Arsenicals / Q000494:pharmacology*; D001665:Binding Sites / Q000502:physiology; D002540:Cerebral Cortex / Q000166:cytology; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism; D009447:Neuroblastoma; D009474:Neurons / Q000166:cytology / Q000201:enzymology*; D017239:Paclitaxel / Q000494:pharmacology; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D051381:Rats; D012694:Serine; D014407:Tumor Cells, Cultured; D016875:tau Proteins / Q000737:chemistry / Q000378:metabolism*
D000972:Antineoplastic Agents, Phytogenic; D001152:Arsenicals; D004791:Enzyme Inhibitors; D016875:tau Proteins; C029341:oxophenylarsine; D012694:Serine; C105547:MARK1 protein, human; C484552:Mark1 protein, rat; D017346:Protein Serine-Threonine Kinases; D017239:Paclitaxel