The distribution and characterization of endogenous protein arginine N-methyltransferase 8 in mouse CNS.
Protein arginine N-methyltransferase (PRMT) 8 was first discovered from a database search for genes harboring four conserved methyltransferase motifs, which shares more than 80% homology to PRMT1 in amino acid [Lee J, Sayegh J, Daniel J, Clarke S, Bedford MT (2005) PRMT8, a new membrane-bound tissue-specific member of the protein arginine methyltransferase family. J Biol Chem 280:32890-32896]. Interestingly, its tissue distribution is strikingly restricted to mouse CNS. To characterize the function in the CNS neurons, we raised an antiserum against PRMT8 to perform immunohistochemistry (IHC) and Western blot analysis. By IHC, the immunoreactivity of endogenous PRMT8 was broadly distributed in the CNS neurons with markedly intense signals in the cerebellum, hippocampal formation, and cortex, but was not detected in the cerebellar granular layer. In some subset of the neurons, the immunoreactivity was observed in the dendrites and axon bundles. The subcellular localization of the immunoreactivity was dominantly nuclear, arguing against the original report that exogenously expressed PRMT8 localizes to the plasma membrane via the N-terminal myristoylation. A series of the exogenously expressed proteins with different in-frame translation initiation codons was tested for comparison with the endogenous protein in molecular size. The third initiator codon produced the protein that was equivalent in size to the endogenous and showed a similar localizing pattern in PC12 cells. In conclusion, PRMT8 is a neuron-specific nuclear enzyme and the N-terminus does not contain the glycine end for myristoylation target.
Kousaka
A
A
Department of Anatomy and Neuroscience, Graduate School of Medicine, The Osaka-Hamamatsu Joint Research Center for Child Mental Development, Osaka University, 2-2 Yamadaoka, Suita City, Osaka, Japan.
Mori
Y
Y
Koyama
Y
Y
Taneda
T
T
Miyata
S
S
Tohyama
M
M
D000818:Animals; D001369:Axons / Q000201:enzymology / Q000378:metabolism; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism; D002467:Cell Nucleus / Q000201:enzymology / Q000378:metabolism; D018387:Codon, Initiator; D003712:Dendrites / Q000201:enzymology / Q000378:metabolism; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009474:Neurons / Q000201:enzymology / Q000378:metabolism*; D016716:PC12 Cells; D011484:Protein-Arginine N-Methyltransferases / Q000235:genetics / Q000378:metabolism*; D051381:Rats
D018387:Codon, Initiator; C546992:PRMT8 protein, mouse; D011484:Protein-Arginine N-Methyltransferases