This study demonstrates a role for S100A10 in autophagy. Silencing of S100A10 reduces IFN-γ-induced autophagosome formation. S100A10 interacts and regulates ULK1 localization to endoplasmic reticulum–mitochondria contact sites. S100A10 regulates autophagy-dependent high mobility group protein B1 release. Our study reveals a regulatory role for S100A10 on ULK1 in IFN-γ-induced autophagy.