PT  - JOURNAL ARTICLE
AU  - SJ Pleasure
AU  - VM Lee
AU  - DL Nelson
TI  - Site-specific phosphorylation of the middle molecular weight human neurofilament protein in transfected non-neuronal cells
AID  - 10.1523/JNEUROSCI.10-07-02428.1990
DP  - 1990 Jul 01
TA  - The Journal of Neuroscience
PG  - 2428--2437
VI  - 10
IP  - 7
4099  - http://www.jneurosci.org/content/10/7/2428.short
4100  - http://www.jneurosci.org/content/10/7/2428.full
SO  - J. Neurosci.1990 Jul 01; 10
AB  - We expressed the human midsized neurofilament subunit (NF-M) using genomic DNA in mouse L cells and showed that it is transcribed and translated into a protein capable of assembly into the cytoskeleton and of forming a filamentous network that colocalizes with the endogenous vimentin filaments. Moreover, human NF-M expressed in L cells is phosphorylated at sites within the multiphosphorylation repeat (MPR), i.e., the major sites of phosphorylation of NF-M in vivo. We also expressed a genomic construct lacking the MPR domain in the native molecule and showed that this MPR(-) protein also was expressed and formed a filamentous network despite diminished incorporation of radiolabeled phosphate. Two major conclusions emerged from the work described in this paper: human NF-M is translated, assembled, and phosphorylated at physiological sites without the need of any other specific neuronal proteins; phosphorylation sites other than the MPR are present within NF-M which may play a role in synthesis, assembly, and degradation of NF protein in humans.