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A novel dRYBP–SCF complex functions to inhibit apoptosis in Drosophila

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Abstract

A balanced response to intrinsic and extrinsic apoptotic signals is crucial to support homeostatic development and animal survival. Regulation of activation and inhibition of apoptotic pathways involves diverse mechanisms including protein ubiquitylation to control expression levels of apoptotic factors. Here we report that drosophila Ring and YY1 Binding Protein (dRYBP) protein interacts both genetically and biochemically with the E3 ubiquitin ligase SKPA, dCULLIN, F-box (SCF) complex to synergistically inhibit apoptosis in Drosophila. Further, we show that the loss of skpA function activates the intrinsic pathway of apoptosis and down-regulates the levels of expression of the anti-apoptotic DIAP1 protein. Accordingly, the apoptosis induced by inactivation of skpA and dRYBP is rescued by loss of function of the pro-apoptotic gene reaper and overexpression of DIAP1. Of interest, we also find that high levels of SKPA protein rescue the wing phenotype induced by overexpression of Reaper protein. Finally, we demonstrate that overexpression of SKPA inhibits both developmental and radiation-induced apoptosis. We propose that the function of the dRYBP–SCF complex in the inhibition of apoptosis might possibly be to control the levels of the pro-apoptotic and anti-apoptotic proteins most likely by promoting their ubiquitylation and consequently, proteasomal degradation. Given the evolutionary conservation of the dRYBP and the SCF proteins, our results suggest that their mammalian homologs may function in balancing cell survival versus cell death during normal and pathological development.

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Acknowledgments

We thank our colleges Carolina Simoes da Silva and Ricardo Aparicio for fruitful discussions and Keith Harshman for carefully reading the manuscript. We are indebted to Peter C. Verrijzer and members of his laboratory for sharing the mass spectrometry data. We thank the Consolider transgenic Drosophila facility for transgenic flies; the Bloomington Stock Center, the TRiP at Harvard Medical School (NIH/NIGMS R01-GM084947) and the Vienna Drosophila RNAi Center for providing stocks; James Wakefield for the anti-SKPA antibody; Herman Steller for the anti-DIAP1 antibody and the UAS-rpr-HA transgenic flies; Peter C. Verrijzer for embryonic nuclear extracts and Pat H. O’Farrell for the UAS-skpA construct. This work was supported by grants from the Dirección General de Investigación (BFU2008-01154) to A.B, the Consolider Ingenio 2010 Program of the Ministerio de Ciencia e Innovación (CSD 2007-00008) to A.B., by an institutional grant to the Centro de Biología Molecular Severo Ochoa from the Fundación Ramón Areces.

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The authors declare that they have no conflict of interest.

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  1. Centro de Biología Molecular “Severo Ochoa” CSIC-UAM, c/Nicolás Cabrera 1, 28049, Madrid, Spain

    Sol Fereres, Rocío Simón & Ana Busturia

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  1. Sol Fereres
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  2. Rocío Simón
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Correspondence to Ana Busturia.

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Fereres, S., Simón, R. & Busturia, A. A novel dRYBP–SCF complex functions to inhibit apoptosis in Drosophila . Apoptosis 18, 1500–1512 (2013). https://doi.org/10.1007/s10495-013-0897-4

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