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Article Dans Une Revue Molecular Vision Année : 2003

A proline-rich domain in the gamma subunit of phosphodiesterase 6 mediates interaction with SH3-containing proteins

Fabrice Morin
  • Fonction : Auteur
Brigitte Vannier
Florent Houdart
  • Fonction : Auteur
Matthieu Régnacq
  • Fonction : Auteur
  • PersonId : 870169
  • IdRef : 12701652X
Pierre Voisin
  • Fonction : Auteur

Résumé

Purpose: Phosphodiesterase 6 (PDE6) is the primary effector of phototransduction in vertebrate photoreceptors. Previous studies described the expression of the regulatory subunit of rod PDE6 (Pγ-rod) in non-photosensitive tissues of the adult rat and the effects of this protein on MAP kinase pathways. Upon examination of the Pγ-rod sequence, we detected a proline-rich domain that might reveal its ability to interact with SH3-containing proteins. Therefore, the present study was initiated to identify new protein partners of Pγ-rod. Methods: A yeast two-hybrid screen of a rat brain cDNA library was performed using Pγ-rod as a bait. Pγ-rod-SH3 interaction was confirmed by GST pull-down of in vitro-translated proteins. The aminoacids involved in the interaction were mapped by site-directed mutagenesis. Rnase protection assay, RT-PCR and western blot analysis were used to detect Pγ-rod expression in various rat tissues. Results: A clone was isolated twice, that consisted essentially of the SH3 domain of the formin-binding protein 17 (FBP17). This interaction was confirmed by GST pull-down. Mutational analysis of the Pγ-rod-FBP17 interaction confirmed it involved the proline-rich domain of Pγ-rod and the SH3 domain of FBP17. This proline-rich domain also allowed Pγ-rod to interact with Cdc42-interacting protein 4 (CIP4), another SH3-containing protein. RT-PCR and Rnase protection assay detected different amounts of Pγ-rod mRNA in adult and embryonic rat tissues. Western blots confirmed the presence of low levels of Pγ-rod protein only in embryonic tissues. Conclusions: Our data suggest that Pγ-rod participates in SH3-mediated cellular pathways and may therefore play a wider role than previously appreciated. One possibility is that FBP17 interaction with sorting nexin 2 might connect Pγ- rod to receptor tyrosine kinase recycling. However, further studies are still required to identify the diversity of SH3- containing proteins that interact with Pγ-rod. This effort should provide a rationale to understand how Pγ-rod can affect receptor internalization-dependent MAP kinase activity.
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Dates et versions

hal-00105585 , version 1 (11-10-2006)

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  • HAL Id : hal-00105585 , version 1

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Fabrice Morin, Brigitte Vannier, Florent Houdart, Matthieu Régnacq, Thierry Bergès, et al.. A proline-rich domain in the gamma subunit of phosphodiesterase 6 mediates interaction with SH3-containing proteins. Molecular Vision, 2003, 9, pp.449-459. ⟨hal-00105585⟩

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