Systematic Analysis of Human Protein Complexes Identifies Chromosome Segregation Proteins

• DOI: 10.1126/science.1181348
• Publication Date: 2010-04-02T02:27:57Z
• Authors: James R. A. Hutchins; Yusuke Toyoda; Björn Hegemann; Ina Poser; Jean-Karim Hériché; Martina M. Sykora; Martina Augsburg; Otto Hudecz; Bettina A. Buschhorn; Jutta Bulkescher; Christian Conrad; David Comartin; Alexander Schleiffer; Mihail Sarov; Andrei Pozniakovsky; Mikolaj Michal Sl...; Siegfried Schlois...; Ines Steinmacher; Marit Leuschner; Andrea Ssykor; Steffen Lawo; Laurence Pelletier; Holger Stark; Kim Nasmyth; Jan Ellenberg; Richard Durbin; Frank Buchholz; Karl Mechtler; Anthony A. Hyman; Jan-Michael Peters
• Keywords: Centrosome; Chromosomes, Artificial, Bacterial; Green Fluorescent Proteins; Mitosis; Ubiquitin-Protein Ligase Complexes; Genomics; Spindle Apparatus; Anaphase-Promoting Complex-Cyclosome; Open Reading Frames; Protein Subunits; Tubulin; Chromosome Segregation; Multiprotein Complexes; Databases, Genetic; Protein Interaction Mapping; Humans; RNA Interference; HeLa Cells; Protein Binding

Chromosome segregation and cell division are essential, highly ordered processes that depend on numerous protein complexes. Results from recent RNA interference screens indicate the identity composition of these complexes is incompletely understood. Using gene tagging bacterial artificial chromosomes, localization, tandem-affinity purification-mass spectrometry, MitoCheck consortium has analyzed about 100 human complexes, many which had not or only been characterized. This work led to discovery previously unknown, evolutionarily conserved subunits anaphase-promoting complex gamma-tubulin ring complex--large essential for spindle assembly chromosome segregation. The approaches we describe here generally applicable high-throughput follow-up analyses phenotypic in mammalian cells.