Fig. 1: MitoID identifies ARMH3 as a potential ARL5 effector.
From: ARMH3 is an ARL5 effector that promotes PI4KB-catalyzed PI4P synthesis at the trans-Golgi network
a Schematic representation in N- to C-terminal direction of control and Mito-ARL5 constructs used in MitoID. Constructs comprise the mitochondrial targeting sequence (MTS) from TOM20, followed by the biotin ligase BioID2, with or without constitutively active (Q70L) or inactive (T30N) ARL5A or ARL5B lacking the 13-amino-acid N-terminal α-helix. These constructs were expressed by transient transfection in HEK293T cells. b Schematic representation of the MitoID procedure. The Mito-ARL5 or control constructs are targeted to mitochondria, and neighboring proteins biotinylated, affinity-purified, and identified by mass spectrometry. Schematic partially created in BioRender. Bonifacino, J. (2024) BioRender.com/w68r614. c Graphs showing the log2(fold-change) of proteins identified in Mito-ARL5 relative to control datasets vs the -log10(p-value). Select hits are labeled in color. ARMH3, VPS52, and VPS13C were identified in the Q70L but not T30N forms of ARL5A and/or ARL5B, and were thus deemed of interest. In contrast, ACACA, FKBP8, HLTF, ILKAP, MFF, and WDR48 were identified in both the Q70L and T30N forms between ARL5 variants and were not further investigated. Source data are provided in the Source Data file.
