Rab11-Rab8 cascade dynamics in primary cilia and membrane tubules

The Rab11-Rab8 cascade mediated by the Rab8 guanine nucleotide exchange factor (GEF), Rabin8, orchestrates multiple membrane transport processes, but Rab membrane loading and exchange dynamics are unclear. Here, we use advanced fluorescence imaging approaches to characterize Rab11, Rab8, and Rabin8 protein dynamics. Using fluorescence ablation and recovery studies (FRAP), we show that Rab8 ciliary trafficking requires Rab11 and Rabin8. Reciprocally, we discover that Rab11 is recruited to cilia during ciliogenesis in association with Rab8. We uncover a requirement for this cascade in Rab8 association with long tubular membranes (LTMs) in human cells and zebrafish embryos. Membrane exchange dynamics of Rab11 on Rab8 LTMs is shown using super-resolution imaging, along with a dependency on Rabin8 GEF activity. Finally, cascade-dependent Rab8 loading onto enlarged Rab11-Rabin8 membrane structures is discussed. This study demonstrates that the Rab11-Rab8 cascade involves membrane conversion and expands our understanding of the cellular multifunctionality of this trafficking pathway.