Author/Authors :
Shinya Ohta، نويسنده , , Jimi-Carlo Bukowski-Wills، نويسنده , , Luis Sanchez-Pulido، نويسنده , , Flavia de Lima Alves، نويسنده , , Laura Wood، نويسنده , , Zhuo A. Chen، نويسنده , , Melpi Platani، نويسنده , , Lutz Fischer، نويسنده , , Damien F. Hudson، نويسنده , , Chris P. Ponting، نويسنده , , Tatsuo Fukagawa، نويسنده , , William C. Earnshaw، نويسنده , , Juri Rappsilber، نويسنده ,
Abstract :
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 ∼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 ∼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.
