Systematic analysis of human protein complexes identifies chromosome segregation proteins.
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.
Hutchins
James R A
JR
Research Institute of Molecular Pathology (IMP), Dr. Bohr-Gasse 7, A-1030 Vienna, Austria.
Toyoda
Yusuke
Y
Hegemann
Björn
B
Poser
Ina
I
Hériché
Jean-Karim
JK
Sykora
Martina M
MM
Augsburg
Martina
M
Hudecz
Otto
O
Buschhorn
Bettina A
BA
Bulkescher
Jutta
J
Conrad
Christian
C
Comartin
David
D
Schleiffer
Alexander
A
Sarov
Mihail
M
Pozniakovsky
Andrei
A
Slabicki
Mikolaj Michal
MM
Schloissnig
Siegfried
S
Steinmacher
Ines
I
Leuschner
Marit
M
Ssykor
Andrea
A
Lawo
Steffen
S
Pelletier
Laurence
L
Stark
Holger
H
Nasmyth
Kim
K
Ellenberg
Jan
J
Durbin
Richard
R
Buchholz
Frank
F
Mechtler
Karl
K
Hyman
Anthony A
AA
Peters
Jan-Michael
JM
D064173:Anaphase-Promoting Complex-Cyclosome; D018385:Centrosome / Q000378:metabolism; D020090:Chromosome Segregation; D022202:Chromosomes, Artificial, Bacterial; D030541:Databases, Genetic; D023281:Genomics; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D008938:Mitosis; D046912:Multiprotein Complexes / Q000378:metabolism*; D016366:Open Reading Frames; D011485:Protein Binding; D025941:Protein Interaction Mapping; D021122:Protein Subunits / Q000378:metabolism; D034622:RNA Interference; D008941:Spindle Apparatus / Q000378:metabolism*; D014404:Tubulin / Q000378:metabolism*; D043743:Ubiquitin-Protein Ligase Complexes / Q000378:metabolism*
D046912:Multiprotein Complexes; D021122:Protein Subunits; D014404:Tubulin; D049452:Green Fluorescent Proteins; D043743:Ubiquitin-Protein Ligase Complexes; D064173:Anaphase-Promoting Complex-Cyclosome