The sterol-regulating human ARV1 binds cholesterol and phospholipids through its conserved ARV1 homology domain.

Evidence suggests that ARV1 regulates sterol movement within the cell. Saccharomyces cerevisiae cells lacking ScArv1 have defects in sterol trafficking, distribution, and biosynthesis. HepG2 cells treated with hARV1 antisense oligonucleotides accumulate cholesterol in the endoplasmic reticulum. Mice lacking Arv1 have a lean phenotype when fed a high fat diet and show no signs of liver triglyceride or cholesterol accumulation, suggesting a role for Arv1 in lipid transport. Here, we explored the direct lipid-binding activity of recombinant human ARV1 using in vitro lipid-binding assays. ARV1 lipid-binding activity was observed within the first N-terminal 98 amino acids containing the conserved ARV1 homology domain (AHD). The zinc-binding domain and conserved cysteine clusters within the AHD were necessary for lipid binding. Both full-length ARV1 and the AHD bound cholesterol, several phospholipids, and phosphoinositides with high affinity. The AHD showed the highest binding affinity for monophosphorylated phosphoinositides. Several conserved amino acids within the AHD were necessary for phospholipid binding. Biochemical studies suggested that ARV1 exists as a dimer in cells, with oligomerization being critical for ARV1 function, as amino acid mutations predicted to have a negative effect on dimerization caused weakened or complete loss of lipid binding. Our results show for the first time that human ARV1 can directly bind cholesterol and phospholipids. How this activity may function to regulate lipid binding and maintain proper lipid trafficking and/or transport in cells requires further studies.