Transducin β-Subunit Can Interact with Multiple G-Protein γ-Subunits to Enable Light Detection by Rod Photoreceptors

The heterotrimeric G-protein transducin mediates visual signaling in vertebrate photoreceptor cells. Many aspects of the function of transducin were learned from knock-out mice lacking its individual subunits. Of particular interest is the knockout of its rod-specific γ-subunit (Gγ₁). Two studies using independently generated mice documented that this knockout results in a considerable >60-fold reduction in the light sensitivity of affected rods, but provided different interpretations of how the remaining α-subunit (Gα_t) mediates phototransduction without its cognate Gβ₁γ₁-subunit partner. One study found that the light sensitivity reduction matched a corresponding reduction in Gα_t content in the light-sensing rod outer segments and proposed that Gα_t activation is supported by remaining Gβ₁ associating with other Gγ subunits naturally expressed in photoreceptors. In contrast, the second study reported the same light sensitivity loss but a much lower, only approximately sixfold, reduction of Gα_t and proposed that the light responses of these rods do not require Gβγ at all. To resolve this controversy and elucidate the mechanism driving visual signaling in Gγ₁ knock-out rods, we analyzed both mouse lines side by side. We first determined that the outer segments of both mice have identical Gα_t content, which is reduced ∼65-fold from the wild-type (WT) level. We further demonstrated that the remaining Gβ₁ is present in a complex with endogenous Gγ₂ and Gγ₃ subunits and that these complexes exist in wild-type rods as well. Together, these results argue against the idea that Gα_t alone supports light responses of Gγ₁ knock-out rods and suggest that Gβ₁γ₁ is not unique in its ability to mediate vertebrate phototransduction.