ausblenden:
Schlagwörter:
Anaphase-Promoting Complex-Cyclosome
Centrosome/metabolism
*Chromosome Segregation
Chromosomes, Artificial, Bacterial
Databases, Genetic
Genomics
Green Fluorescent Proteins
HeLa Cells
Humans
*Mitosis
Multiprotein Complexes/*metabolism
Open Reading Frames
Protein Binding
Protein Interaction Mapping
Protein Subunits/metabolism
RNA Interference
Spindle Apparatus/*metabolism
Tubulin/*metabolism
Ubiquitin-Protein Ligase Complexes/*metabolism
Zusammenfassung:
Chromosome segregation and cell division are essential, highly ordered processes that depend on numerous protein complexes. Results from recent RNA interference screens indicate that the identity and composition of these protein complexes is incompletely understood. Using gene tagging on bacterial artificial chromosomes, protein localization, and tandem-affinity purification-mass spectrometry, the MitoCheck consortium has analyzed about 100 human protein complexes, many of which had not or had only incompletely been characterized. This work has led to the discovery of previously unknown, evolutionarily conserved subunits of the anaphase-promoting complex and the gamma-tubulin ring complex--large complexes that are essential for spindle assembly and chromosome segregation. The approaches we describe here are generally applicable to high-throughput follow-up analyses of phenotypic screens in mammalian cells.
