Rab Interacting Molecules 2 and 3 Directly Interact with the Pore-Forming Ca_V1.3 Ca²⁺ Channel Subunit and Promote Its...

Rab interacting molecules (RIMs) are multi-domain proteins that positively regulate the number of Ca²⁺ channels at the presynaptic active zone (AZ). Several molecular mechanisms have been demonstrated for RIM-binding to components of the presynaptic Ca²⁺ channel complex, the key signaling element at the AZ. Here, we report an interaction of the C₂B domain of RIM2α and RIM3γ with the C-terminus of the pore-forming α-subunit of Ca_V1.3 channels (Ca_V1.3α1), which mediate stimulus-secretion coupling at the ribbon synapses of cochlear inner hair cells (IHCs). Co-expressing full-length RIM2α with a Ca²⁺ channel complex closely resembling that of IHCs (Ca_V1.3α1-Ca_Vß2a) in HEK293 cells doubled the Ca²⁺-current and shifted the voltage-dependence of Ca²⁺ channel activation by approximately +3 mV. Co-expression of the short RIM isoform RIM3γ increased the Ca_V1.3α1-Ca_Vß2a-mediated Ca²⁺-influx in HEK293 cells, but disruption of RIM3γ in mice left Ca²⁺-influx in IHCs and hearing intact. In conclusion, we propose that RIM2α and RIM3γ directly interact with the C-terminus of the pore-forming subunit of Ca_V1.3 Ca²⁺ channels and positively regulate their plasma membrane expression in HEK293 cells.