A protein interaction map of the mitotic spindle.
The mitotic spindle consists of a complex network of proteins that segregates chromosomes in eukaryotes. To strengthen our understanding of the molecular composition, organization, and regulation of the mitotic spindle, we performed a system-wide two-hybrid screen on 94 proteins implicated in spindle function in Saccharomyces cerevisiae. We report 604 predominantly novel interactions that were detected in multiple screens, involving 303 distinct prey proteins. We uncovered a pattern of extensive interactions between spindle proteins reflecting the intricate organization of the spindle. Furthermore, we observed novel connections between kinetochore complexes and chromatin-modifying proteins and used phosphorylation site mutants of NDC80/TID3 to gain insights into possible phospho-regulation mechanisms. We also present analyses of She1p, a novel spindle protein that interacts with the Dam1 kinetochore/spindle complex. The wealth of protein interactions presented here highlights the extent to which mitotic spindle protein functions and regulation are integrated with each other and with other cellular activities.
Wong
Jonathan
J
Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA.
Nakajima
Yuko
Y
Westermann
Stefan
S
Shang
Ching
C
Kang
Jung-Seog
JS
Goodner
Crystal
C
Houshmand
Pantea
P
Fields
Stanley
S
Chan
Clarence S M
CS
Drubin
David
D
Barnes
Georjana
G
Hazbun
Tony
T
D002843:Chromatin / Q000378:metabolism; D030562:Databases, Protein; D018386:Kinetochores / Q000378:metabolism; D046912:Multiprotein Complexes / Q000378:metabolism; D050505:Mutant Proteins / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D025941:Protein Interaction Mapping; D021122:Protein Subunits / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D008941:Spindle Apparatus / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
D002843:Chromatin; D046912:Multiprotein Complexes; D050505:Mutant Proteins; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins