MSAP is a novel MIR-interacting protein that enhances neurite outgrowth and increases myosin regulatory light chain.
Dynamic interactions between the actin cytoskeleton and specific proteins are crucial for changes in cell shape and motility. Here we describe a novel protein MSAP (MIR-interacting saposin-like protein) that is a positive regulator of neurite outgrowth. MSAP is expressed in different tissues, including brain, and has an apparent molecular weight of 21 kDa. MSAP interacts with the ezrin-radixin-moesin (ERM)-like myosin regulatory light chain-interacting protein (MIR), and the two proteins are co-localized in cell lines and in primary neurons. Overexpression of MSAP enhances neurite out-growth in neuroblastoma and PC12 cells, whereas down-regulation of MSAP using RNA silencing led to inhibition of neurite formation. The stimulation of neurite outgrowth by MSAP was abrogated by the overexpression of MIR, which induced a decrease in the levels of myosin regulatory light chain (MRLC). This reduction in MRLC by MIR was inhibited by blocking the activity of proteasome and by overexpression of MSAP, suggesting an effect on protein stability. Evidence was obtained that MIR decreases MRLC by inducing its ubiquitination. The results show that the levels of MRLC are controlled by MIR via ubiquitination and that the effect of MIR on MRLC is counteracted in the presence of MSAP. MSAP can stabilize MRLC and thus bring about an increase in neurite outgrowth.
Bornhauser
Beat C
BC
Department of Neuroscience and Neurobiology, Uppsala University, Biomedical Center, Box 587, S-75123 Uppsala, Sweden.
Olsson
Per-Anders
PA
Lindholm
Dan
D
D048868:Adaptor Proteins, Signal Transducing; D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000196:embryology / Q000378:metabolism*; D019556:COS Cells; D002352:Carrier Proteins / Q000378:metabolism*; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D005333:Fetus; D006367:HeLa Cells; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D018994:Myosin Light Chains / Q000378:metabolism*; D016501:Neurites / Q000502:physiology*; D010446:Peptide Fragments / Q000737:chemistry; D011994:Recombinant Proteins / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014162:Transfection; D044767:Ubiquitin-Protein Ligases
D048868:Adaptor Proteins, Signal Transducing; C502333:CNPY2 protein, human; D002352:Carrier Proteins; D018994:Myosin Light Chains; D010446:Peptide Fragments; D011994:Recombinant Proteins; C403126:MYLIP protein, human; D044767:Ubiquitin-Protein Ligases