A conventional PKC critical for both the light-dependent and the light-independent regulation of the actin cytoskeleton in Drosophila photoreceptors

Shieh, Bih‐Hwa; Sun, Wesley; Ferng, Darwin

doi:10.1016/j.jbc.2023.104822

Cited by 2 publications

(1 citation statement)

References 52 publications

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“…Green light may evoke a distinct conformation of Rh1 different from that of the blue light. We explored the endocytic mechanism of Rh1 by green light in transgenic flies expressing Rh1-mC [40].…”

Section: Green Light Triggers the Endocytosis Of Rh1-mc In Both Adult...mentioning

confidence: 99%

Differential activation of rhodopsin triggers distinct endocytic trafficking and recycling in vivo via differential phosphorylation

Ferng,

Sun,

Shieh

2024

PLoS ONE

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Activated GPCRs are phosphorylated and internalized mostly via clathrin-mediated endocytosis (CME), which are then sorted for recycling or degradation. We investigated how differential activation of the same GPCR affects its endocytic trafficking in vivo using rhodopsin as a model in pupal photoreceptors of flies expressing mCherry-tagged rhodopsin 1 (Rh1-mC) or GFP-tagged arrestin 1 (Arr1-GFP). Upon blue light stimulation, activated Rh1 recruited Arr1-GFP to the rhabdomere, which became co-internalized and accumulated in cytoplasmic vesicles of photoreceptors. This internalization was eliminated in shits1 mutants affecting dynamin. Moreover, it was blocked by either rdgA or rdgB mutations affecting the PIP2 biosynthesis. Together, the blue light-initiated internalization of Rh1 and Arr1 belongs to CME. Green light stimulation also triggered the internalization and accumulation of activated Rh1-mC in the cytoplasm but with faster kinetics. Importantly, Arr1-GFP was also recruited to the rhabdomere but not co-internalized with Rh1-mC. This endocytosis was not affected in shits1 nor rdgA mutants, indicating it is not CME. We explored the fate of internalized Rh1-mC following CME and observed it remained in cytoplasmic vesicles following 30 min of dark adaptation. In contrast, in the non-CME Rh1-mC appeared readily recycled back to the rhabdomere within five min of dark treatment. This faster recycling may be regulated by rhodopsin phosphatase, RdgC. Together, we demonstrate two distinct endocytic and recycling mechanisms of Rh1 via two light stimulations. It appears that each stimulation triggers a distinct conformation leading to different phosphorylation patterns of Rh1 capable of recruiting Arr1 to rhabdomeres. However, a more stable interaction leads to the co-internalization of Arr1 that orchestrates CME. A stronger Arr1 association appears to impede the recycling of the phosphorylated Rh1 by preventing the recruitment of RdgC. We conclude that conformations of activated rhodopsin determine the downstream outputs upon phosphorylation that confers differential protein-protein interactions.

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Section: Green Light Triggers the Endocytosis Of Rh1-mc In Both Adult...mentioning

confidence: 99%

Differential activation of rhodopsin triggers distinct endocytic trafficking and recycling in vivo via differential phosphorylation

Ferng,

Sun,

Shieh

2024

PLoS ONE

Self Cite

View full text Add to dashboard Cite

show abstract

Differential activation of rhodopsin triggers distinct endocytic trafficking and recycling in vivo via differential phosphorylation

Ferng,

Sun,

Shieh

2024

Preprint

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Activated GPCRs are phosphorylated and internalized mostly via clathrin-mediated endocytosis (CME), which are then sorted for recycling or degradation. We investigated how differential activation of the same GPCR affects its endocytic traffickingin vivousing rhodopsin as a model in flies expressing mCherry-tagged rhodopsin 1 (Rh1-mC) or GFP-tagged arrestin 1 (Arr1-GFP). Upon blue light stimulation, activated Rh1 recruited Arr1-GFP to the rhabdomere, which became co-internalized and accumulated in cytoplasmic vesicles of photoreceptors. This internalization was eliminated inshits1mutants affecting dynamin. Moreover, it was blocked by eitherrdgAorrdgBmutations affecting the PIP2biosynthesis. Together, the blue light-initiated internalization of Rh1 and Arr1 belongs to CME. Green light stimulation also triggered the internalization and accumulation of activated Rh1-mC in the cytoplasm but with faster kinetics. Importantly, Arr1-GFP was also recruited to the rhabdomere but not co-internalized with Rh1-mC. This endocytosis was not affected inshits1norrdgAmutants, indicating it is not CME. We explored the fate of internalized Rh1-mC following CME and observed it remained in cytoplasmic vesicles following 30 min of dark adaptation. In contrast, in the non-CME Rh1-mC appeared readily recycled back to the rhabdomere within five min of dark treatment. This faster recycling may be regulated by rhodopsin phosphatase, RdgC. Together, we demonstrate two distinct endocytic and recycling mechanisms of Rh1 via two light stimulations. It appears that each stimulation triggers a distinct conformation leading to different phosphorylation patterns of Rh1 capable of recruiting Arr1 to rhabdomeres. However, a more stable interaction leads to the co-internalization of Arr1 that orchestrates CME. A stronger Arr1 association appears to impede the recycling of the phosphorylated Rh1 by preventing the recruitment of RdgC. We conclude that conformations of activated rhodopsin determine the downstream outputs upon phosphorylation that confers differential protein-protein interactions.

show abstract

A conventional PKC critical for both the light-dependent and the light-independent regulation of the actin cytoskeleton in Drosophila photoreceptors

Cited by 2 publications

References 52 publications

Differential activation of rhodopsin triggers distinct endocytic trafficking and recycling in vivo via differential phosphorylation

Differential activation of rhodopsin triggers distinct endocytic trafficking and recycling in vivo via differential phosphorylation

Differential activation of rhodopsin triggers distinct endocytic trafficking and recycling in vivo via differential phosphorylation

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