The yeast Ski complex is a hetero-tetramer.
The yeast Ski complex assists the exosome in the degradation of mRNA. The Ski complex consists of three components; Ski2, Ski3, and Ski8, believed to be present in a 1:1:1 stoichiometry. Measuring the mass of intact isolated endogenously expressed Ski complexes by native mass spectrometry we unambiguously demonstrate that the Ski complex has a hetero-tetrameric stoichiometry consisting of one copy of Ski2 and Ski3 and two copies of Ski8. To validate the stoichiometry of the Ski complex, we performed tandem mass spectrometry. In these experiments one Ski8 subunit was ejected concomitant with the formation of a Ski2/Ski3/Ski8 fragment, confirming the proposed stoichiometry. To probe the topology of the Ski complex we disrupted the complex and mass analyzed the thus formed subcomplexes, detecting Ski8-Ski8, Ski2-Ski3, Ski8-Ski2, and Ski8-Ski8-Ski2. Combining all data we construct an improved structural model of the Ski complex.
Department of Biomolecular Mass Spectrometry, Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, The Netherlands.
Silvia A
SA
Synowsky
Albert J R
AJ
Heck
D013058:Mass Spectrometry; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics; D011487:Protein Conformation; D012331:RNA, Fungal / Q000235:genetics*; D012333:RNA, Messenger / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics
D009687:Nuclear Proteins; D012331:RNA, Fungal; D012333:RNA, Messenger; C081929:SKI2 protein, S cerevisiae; C406671:SKI3 protein, S cerevisiae; C079882:SKI8 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins