Unraveling the intricate cargo-BBSome coupling mechanism at the ciliary tip

• Published in: Proceedings of the National Academy of Sciences - PNAS 2023-03, Vol.120 (13), p.e2218819120-e2218819120
• Main Authors: Liu, Yan-Xia, Li, Wen-Juan, Zhang, Rui-Kai, Sun, Sheng-Nan, Fan, Zhen-Chuan
• Format: Article
• Language: English
• Subjects: Adaptor proteins; Bardet-Biedl Syndrome - genetics; Biological Sciences; Cargo; Cilia; Cilia - metabolism; Coupling (molecular); Flagella - metabolism; Guanine nucleotide exchange factor; Guanosine Triphosphate - metabolism; Hedgehog protein; Hedgehog Proteins - metabolism; Humans; Membrane Proteins - metabolism; Membranes; Nucleotides; Phospholipase D; Protein Transport - genetics; Proteins; Signaling
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.2218819120

Certain ciliary transmembrane and membrane-tethered signaling proteins migrate from the ciliary tip to base via retrograde intraflagellar transport (IFT), essential for maintaining their ciliary dynamics to enable cells to sense and transduce extracellular stimuli inside the cell. During this process, the BBSome functions as an adaptor between retrograde IFT trains and these signaling protein cargoes. The Arf-like 13 (ARL13) small GTPase resembles ARL6/BBS3 in facilitating these signaling cargoes to couple with the BBSome at the ciliary tip prior to loading onto retrograde IFT trains for transporting towards the ciliary base, while the molecular basis for how this intricate coupling event happens remains elusive. Here, we report that ARL13 only in a GTP-bound form (ARL13 ) anchors to the membrane for diffusing into cilia. Upon entering cilia, ARL13 undergoes GTPase cycle for shuttling between the ciliary membrane (ARL13 ) and matrix (ARL13 ). To achieve this goal, the ciliary membrane-anchored BBS3 binds the ciliary matrix-residing ARL13 to activate the latter as an ARL13 guanine nucleotide exchange factor. At the ciliary tip, ARL13 recruits the ciliary matrix-residing and post-remodeled BBSome as an ARL13 effector to anchor to the ciliary membrane. This makes the BBSome spatiotemporally become available for the ciliary membrane-tethered phospholipase D (PLD) to couple with. Afterward, ARL13 hydrolyzes GTP for releasing the PLD-laden BBSome to load onto retrograde IFT trains. According to this model, hedgehog signaling defects associated with and mutations in humans could be satisfactorily explained, providing us a mechanistic understanding behind BBSome-cargo coupling required for proper ciliary signaling.