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CSB428H1F Article #2: Chuang et al. SARA and Rhodopsin Transport
Introduction
OS discs are renewed whereby rhodopsin-laden vesicles in the OS axonemal cytoplasm fuse with
nascent discs abundant in PI3P and FYVE domain protein SARA interacts with PI3P, rhodopsin and
syntaxin 3 SNARE to regulate this process.
Rhodopsins C’ tail has an address signal to OS and also binds SARA.
FYVE domain is a conserved double zinc finger motif that binds PI3P specifically.
SARA (smad anchor for receptor activation) has FYVE and is located on EE.
SARA interacts with rhodopsin C
In yeast two hybrid and GST pull down assay, rhodopsin C’ interacts with human SARA. Ab for
rhodopsin also co-immuniprecipitated endogenous mouse SARA.
In HEK cells, SARA colocalized with EEA1 and rhodopsin on EE.
SARA is enriched in axonemeal vesicles
In rods, SARA is found in punctate EE in apical cytoplasm of IS and basal portion of proximal
axoneme including the connecting cilium and basal body.
Rhodopsin was found throughout OS.
SARA-PI3P interaction is necessary for rhodopsin delivery to OS
A PI3P probe consisted of double FUVE finger of Hrs that binds PI3P specifically.
In rods, PI3P was found in OS especially proximal portion.
PI3K Vps34 was found only at proximal portion of OS towards base, likely on discs, but not in IS.
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Document Summary

os discs are renewed whereby rhodopsin-laden vesicles in the os axonemal cytoplasm fuse with nascent discs abundant in pi3p and fyve domain protein sara interacts with pi3p, rhodopsin and syntaxin 3 snare to regulate this process. rhodopsin"s c" tail has an address signal to os and also binds sara. Fyve domain is a conserved double zinc finger motif that binds pi3p specifically. Sara (smad anchor for receptor activation) has fyve and is located on ee. In yeast two hybrid and gst pull down assay, rhodopsin c" interacts with human sara. Ab for rhodopsin also co-immuniprecipitated endogenous mouse sara. In hek cells, sara colocalized with eea1 and rhodopsin on ee. In rods, sara is found in punctate ee in apical cytoplasm of is and basal portion of proximal axoneme including the connecting cilium and basal body. Sara-pi3p interaction is necessary for rhodopsin delivery to os.

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