TY - JOUR
T1 - The Protein Composition of Mitotic Chromosomes Determined Using Multiclassifier Combinatorial Proteomics
AU - Ohta, Shinya
AU - Bukowski-Wills, Jimi-Carlo
AU - Sanchez-Pulido, Luis
AU - Alves, Flavia de Lima
AU - Wood, Laura
AU - Chen, Zhuo A.
AU - Platani, Melpi
AU - Fischer, Lutz
AU - Hudson, Damien F.
AU - Ponting, Chris
AU - Fukagawa, Tatsuo
AU - Earnshaw, William C.
AU - Rappsilber, Juri
PY - 2010/9/3
Y1 - 2010/9/3
N2 - Despite many decades of study, mitotic chromosome structure and composition remain poorly characterized. Here, we have integrated quantitative proteomics with bioinformatic analysis to generate a series of independent classifiers that describe the similar to 4,000 proteins identified in isolated mitotic chromosomes. Integrating these classifiers by machine learning uncovers functional relationships between protein complexes in the context of intact chromosomes and reveals which of the similar to 560 uncharacterized proteins identified here merits further study. Indeed, of 34 GFP-tagged predicted chromosomal proteins, 30 were chromosomal, including 13 with centromere-association. Of 16 GFP-tagged predicted nonchromosomal proteins, 14 were confirmed to be nonchromosomal. An unbiased analysis of the whole chromosome proteome from genetic knockouts of kinetochore protein Ska3/Rama1 revealed that the APC/C and RanBP2/RanGAP1 complexes depend on the Ska complex for stable association with chromosomes. Our integrated analysis predicts that up to 97 new centromere-associated proteins remain to be discovered in our data set.
AB - Despite many decades of study, mitotic chromosome structure and composition remain poorly characterized. Here, we have integrated quantitative proteomics with bioinformatic analysis to generate a series of independent classifiers that describe the similar to 4,000 proteins identified in isolated mitotic chromosomes. Integrating these classifiers by machine learning uncovers functional relationships between protein complexes in the context of intact chromosomes and reveals which of the similar to 560 uncharacterized proteins identified here merits further study. Indeed, of 34 GFP-tagged predicted chromosomal proteins, 30 were chromosomal, including 13 with centromere-association. Of 16 GFP-tagged predicted nonchromosomal proteins, 14 were confirmed to be nonchromosomal. An unbiased analysis of the whole chromosome proteome from genetic knockouts of kinetochore protein Ska3/Rama1 revealed that the APC/C and RanBP2/RanGAP1 complexes depend on the Ska complex for stable association with chromosomes. Our integrated analysis predicts that up to 97 new centromere-associated proteins remain to be discovered in our data set.
KW - HUMAN METAPHASE CHROMOSOMES
KW - OUTER KINETOCHORE
KW - QUANTITATIVE PROTEOMICS
KW - CELL-DIVISION
KW - IN-VIVO
KW - CENP-A
KW - COMPLEX
KW - REVEALS
KW - IDENTIFICATION
KW - QUANTIFICATION
UR - http://www.scopus.com/inward/record.url?scp=77956167600&partnerID=8YFLogxK
U2 - 10.1016/j.cell.2010.07.047
DO - 10.1016/j.cell.2010.07.047
M3 - Article
VL - 142
SP - 810
EP - 821
JO - Cell
JF - Cell
SN - 0092-8674
IS - 5
ER -