Functional genomics reveals genes involved in protein secretion and Golgi organization

Yeast genetics and in vitro biochemical analysis have identified numerous genes involved in protein secretion. 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.

[1]  R. Schekman,et al.  Secretion and cell-surface growth are blocked in a temperature-sensitive mutant of Saccharomyces cerevisiae , 1979, Proceedings of the National Academy of Sciences.

[2]  J. Rothman,et al.  A coat subunit of Golgi-derived non-clathrin-coated vesicles with homology to the clathrin-coated vesicle coat protein β-adaptin , 1991, Nature.

[3]  T. Kreis,et al.  Involvement of β-COP in membrane traffic through the Golgi complex , 1991 .

[4]  C. Futter,et al.  Transport into and out of the Golgi complex studied by transfecting cells with cDNAs encoding horseradish peroxidase , 1994, The Journal of cell biology.

[5]  J. Rothman,et al.  Mechanisms of intracellular protein transport , 1994, Nature.

[6]  K. Kaiser,et al.  Deficiency of protein phosphatase 2A uncouples the nuclear and centrosome cycles and prevents attachment of microtubules to the kinetochore in Drosophila microtubule star (mts) embryos. , 1996, Journal of cell science.

[7]  V. Malhotra,et al.  The mechanism of Golgi segregation during mitosis is cell type-specific. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[8]  H. Pelham,et al.  SNAREs and membrane fusion in the Golgi apparatus. , 1998, Biochimica et biophysica acta.

[9]  D. Sharp Cell Division: MAST Sails Through Mitosis , 2002, Current Biology.

[10]  G. Warren,et al.  Golgi architecture and inheritance. , 2002, Annual review of cell and developmental biology.

[11]  N. Heisterkamp,et al.  Interaction of the small GTPase Rac3 with NRBP, a protein with a kinase-homology domain. , 2002, International journal of molecular medicine.

[12]  A. Coulson,et al.  A functional genomic analysis of cell morphology using RNA interference , 2003, Journal of biology.

[13]  F. Uhlmann Separase regulation during mitosis. , 2003, Biochemical Society symposium.

[14]  E. Hartmann,et al.  Use1p is a yeast SNARE protein required for retrograde traffic to the ER , 2003, The EMBO journal.

[15]  Norbert Perrimon,et al.  Parallel Chemical Genetic and Genome-Wide RNAi Screens Identify Cytokinesis Inhibitors and Targets , 2004, PLoS biology.

[16]  V. Chow,et al.  The non-structural 3 (NS3) protein of dengue virus type 2 interacts with human nuclear receptor binding protein and is associated with alterations in membrane structure. , 2004, Virus research.

[17]  N. Perrimon,et al.  Genome-Wide RNAi Analysis of Growth and Viability in Drosophila Cells , 2004, Science.

[18]  N. Perrimon,et al.  Functional Genomic Analysis of the Wnt-Wingless Signaling Pathway , 2005, Science.