Skip to Content
Merck
CN
  • Huntingtin-mediated axonal transport requires arginine methylation by PRMT6.

Huntingtin-mediated axonal transport requires arginine methylation by PRMT6.

Cell reports (2021-04-15)
Alice Migazzi, Chiara Scaramuzzino, Eric N Anderson, Debasmita Tripathy, Ivó H Hernández, Rogan A Grant, Michela Roccuzzo, Laura Tosatto, Amandine Virlogeux, Chiara Zuccato, Andrea Caricasole, Tamara Ratovitski, Christopher A Ross, Udai B Pandey, José J Lucas, Frédéric Saudou, Maria Pennuto, Manuela Basso
ABSTRACT

The huntingtin (HTT) protein transports various organelles, including vesicles containing neurotrophic factors, from embryonic development throughout life. To better understand how HTT mediates axonal transport and why this function is disrupted in Huntington's disease (HD), we study vesicle-associated HTT and find that it is dimethylated at a highly conserved arginine residue (R118) by the protein arginine methyltransferase 6 (PRMT6). Without R118 methylation, HTT associates less with vesicles, anterograde trafficking is diminished, and neuronal death ensues-very similar to what occurs in HD. Inhibiting PRMT6 in HD cells and neurons exacerbates mutant HTT (mHTT) toxicity and impairs axonal trafficking, whereas overexpressing PRMT6 restores axonal transport and neuronal viability, except in the presence of a methylation-defective variant of mHTT. In HD flies, overexpressing PRMT6 rescues axonal defects and eclosion. Arginine methylation thus regulates HTT-mediated vesicular transport along the axon, and increasing HTT methylation could be of therapeutic interest for HD.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
S-(5′-Adenosyl)-L-methionine chloride dihydrochloride, ≥75%
Sigma-Aldrich
Cytosine β-D-arabinofuranoside, crystalline, ≥90% (HPLC)
Sigma-Aldrich
Monoclonal Anti-β-Tubulin I antibody produced in mouse, clone SAP.4G5, ascites fluid
Sigma-Aldrich
Papain from papaya latex, lyophilized powder, ≥10 units/mg protein
Sigma-Aldrich
Trypsin inhibitor from chicken egg white, Type II-O, Partially purified ovomucoid, containing ovoinhibitor
Sigma-Aldrich
Phosphatase Inhibitor Cocktail 2, aqueous solution (dark coloration may develop upon storage, which does not affect the activity)
Sigma-Aldrich
L-Cystine, from non-animal source, meets EP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
Anti-Huntingtin Protein Antibody, a.a. 181-810, clone 1HU-4C8, ascites fluid, clone 1HU-4C8, Chemicon®
Sigma-Aldrich
Anti-MAP2 Antibody, Chemicon®, from chicken
Sigma-Aldrich
Anti-dimethyl-Arginine Antibody, asymmetric (ASYM24), serum, Upstate®
Sigma-Aldrich
Earle′s Balanced Salt Solution 10x, Without sodium bicarbonate, 10 ×, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Poly-D-lysine hydrobromide, average mol wt 30,000-70,000, lyophilized powder, γ-irradiated, BioReagent, suitable for cell culture
Sigma-Aldrich
Deoxyribonuclease I from bovine pancreas, Type IV, lyophilized powder, ≥2,000 Kunitz units/mg protein
Sigma-Aldrich
Adenosine, periodate oxidized, ≥93%
Sigma-Aldrich
L-Glutathione reduced, ≥98.0%
Sigma-Aldrich
Anti-Calnexin antibody produced in rabbit, IgG fraction of antiserum, buffered aqueous solution
Sigma-Aldrich
bisBenzimide H 33342 trihydrochloride, ≥98% (HPLC)
Sigma-Aldrich
Polyethylenimine, branched, average Mw ~25,000 by LS, average Mn ~10,000 by GPC, branched
Sigma-Aldrich
Duolink® In Situ Red Starter Kit Mouse/Rabbit
Sigma-Aldrich
Anti-Green Fluorescent Protein Antibody, original GFP only, clone 264-449-2, clone 264-449-2, Chemicon®, from mouse
Roche
Anti-GFP, from mouse IgG1κ (clones 7.1 and 13.1)