Intermediate filament disassembly in cultured dorsal root ganglion neurons is associated with amino-terminal head domain phosphorylation of specific subunits.
We previously reported that activation of protein kinase A in cultured rat dorsal root ganglion neurons, treated concomitantly with low concentrations of okadaic acid that selectively inhibit protein phosphatase-2A, enhanced the Triton X-100 solubility of neurofilament triplet proteins. We now show that peripherin and alpha-internexin follow the same fragmentation profile as the neurofilament subunits, consistent with the notion that all five cytoplasmic intermediate filament proteins in these neurons form an integrated filamentous network whose assembly can be modulated by protein kinase A. Similar to the situation previously observed for the light neurofilament subunit, there was a strong correlation between phosphorylation of the amino-terminal head domain of peripherin and filament fragmentation. In contrast, insignificant levels of 32P were incorporated into alpha-internexin under conditions promoting disassembly, indicating that phosphorylation of this protein is not involved directly in filament fragmentation. The situation for the mid-sized neurofilament subunit (NFM) was not as clear-cut. Phosphopeptide mapping of NFM revealed many head and tail domain phosphorylation sites. However, changes in NFM head domain phosphorylation under conditions promoting filament disassembly were not as pronounced as for peripherin.
Giasson
B I
BI
Department of Biochemistry, McGill University, Montreal, Quebec, Canada.
Mushynski
W E
WE
D000818:Animals; D002352:Carrier Proteins / Q000502:physiology; D002478:Cells, Cultured; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D004789:Enzyme Activation / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005727:Ganglia, Spinal / Q000166:cytology / Q000502:physiology*; D007381:Intermediate Filament Proteins / Q000502:physiology*; D008562:Membrane Glycoproteins; D008970:Molecular Weight; D009419:Nerve Tissue Proteins / Q000502:physiology; D016900:Neurofilament Proteins / Q000737:chemistry / Q000502:physiology; D009474:Neurons / Q000502:physiology*; D019319:Okadaic Acid / Q000494:pharmacology; D010449:Peptide Mapping; D064531:Peripherins; D010766:Phosphorylation; D051381:Rats
D002352:Carrier Proteins; D004791:Enzyme Inhibitors; D007381:Intermediate Filament Proteins; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins; D064531:Peripherins; C046643:alpha-internexin; D019319:Okadaic Acid; D017868:Cyclic AMP-Dependent Protein Kinases