Abstract
Yeast genetics and in vitro biochemical analysis have identified numerous genes involved in protein secretion1,2. As compared with yeast, however, the metazoan secretory pathway is more complex and many mechanisms that regulate organization of the Golgi apparatus remain poorly characterized. We performed a genome-wide RNA-mediated interference screen in a Drosophila cell line to identify genes required for constitutive protein secretion. We then classified the genes on the basis of the effect of their depletion on organization of the Golgi membranes. Here we show that depletion of class A genes redistributes Golgi membranes into the endoplasmic reticulum, depletion of class B genes leads to Golgi fragmentation, depletion of class C genes leads to aggregation of Golgi membranes, and depletion of class D genes causes no obvious change. Of the 20 new gene products characterized so far, several localize to the Golgi membranes and the endoplasmic reticulum.
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Acknowledgements
We thank members of the Malhotra laboratory for discussions; members of the DRSC for advice; the Institute for Chemistry and Cell Biology for use of their Cybio robot; and J. Feramisco and members of the UCSD Cancer Center imaging facility for help with microscopy. Work in the Malhotra laboratory is supported by NIH grants and a senior investigator award from Sandler's Program for Asthma Research. N.P. is a Howard Hughes investigator.
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Supplementary information
Supplementary Figure 1
Golgi fragmentation and block in HRP secretion are not due to an arrest in mitosis. (PDF 541 kb)
Supplementary Table 1
1133 dsRNAs inducing a significant inhibition of HRP secretion. (PDF 97 kb)
Supplementary Table 2
154 dsRNAs tested in secondary screens and reasons for their removal. (PDF 36 kb)
Supplementary Table 3
130 genes involved in secretion and Golgi membrane organization. (PDF 32 kb)
Supplementary Legends
Text to accompany the above Supplementary Figure and Supplementary Tables. (DOC 40 kb)
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Bard, F., Casano, L., Mallabiabarrena, A. et al. Functional genomics reveals genes involved in protein secretion and Golgi organization. Nature 439, 604–607 (2006). https://doi.org/10.1038/nature04377
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DOI: https://doi.org/10.1038/nature04377
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