Increased phosphorylation of neurofilament subunits in PC12 cells and rat dorsal root ganglion neurons treated with N-Acetyl-Leu-Leu-norleucinal.
Treatment of PC12 cells or dorsal root ganglion neurons with the protease inhibitor, N-Acetyl-Leu-Leu-norleucinal, stimulated phosphorylation of the mid-sized and heavy neurofilament subunits and caused the heavy subunit in the perikarya of dorsal root ganglion neurons to become hyperphosphorylated. The closely related inhibitor, N-Acetyl-Leu-Leu-methioninal, did not produce a similar effect. Okadaic acid increased the phosphorylation state of the heavy neurofilament subunit in PC12 cells in a fashion similar to N-Acetyl-Leu-Leu-norleucinal and the effect of both compounds together was greater than for either one alone. There was no increase in cyclin-dependent kinase 5-immunoprecipitable histone H1 kinase activity in PC12 cells treated with N-Acetyl-Leu-Leu-norleucinal despite the presence of enzyme protein. The present study demonstrates that a protease inhibitor can induce the hyperphosphorylation of neurofilament subunits to a level normally seen only in axons. This suggests that perturbations in intracellular proteolysis may lead to the accumulation of phosphorylated neurofilament epitopes in neuronal perikarya in certain pathological states. The results also show that the carboxy-terminal tail domains of the two largest neurofilament subunits are phosphorylated even when cyclin dependent kinase 5 is inactive, indicating that other neuronal kinases are involved in the phosphorylation of Lys-Ser-Pro repeats.
Jayaraman
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D
Department of Biochemistry, McGill University, Montreal, Quebec, Canada.
Giasson
B I
BI
Mushynski
W E
WE
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D005727:Ganglia, Spinal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007976:Leupeptins / Q000494:pharmacology*; D008969:Molecular Sequence Data; D016900:Neurofilament Proteins / Q000378:metabolism*; D009474:Neurons / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D016716:PC12 Cells; D010766:Phosphorylation / Q000187:drug effects; D011233:Precipitin Tests; D011494:Protein Kinases / Q000378:metabolism; D051381:Rats; D013268:Stimulation, Chemical
D007976:Leupeptins; D016900:Neurofilament Proteins; C063261:acetylleucyl-leucyl-norleucinal; D011494:Protein Kinases