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Related Experiment Video

Updated: Jul 9, 2026

A Modified Yeast-one Hybrid System for Heteromeric Protein Complex-DNA Interaction Studies

A Modified Yeast-one Hybrid System for Heteromeric Protein Complex-DNA Interaction Studies

Published on: July 24, 2017

The yeast Ski complex is a hetero-tetramer.

Silvia A Synowsky¹, Albert J R Heck

¹Department of Biomolecular Mass Spectrometry, Bijvoet Center for Biomolecular Research, Utrecht University, Utrecht, The Netherlands.

Protein Science : a Publication of the Protein Society

|November 29, 2007

Summary

The yeast Ski complex, crucial for mRNA degradation, has a revised stoichiometry. Native mass spectrometry reveals a hetero-tetrameric structure with one Ski2, one Ski3, and two Ski8 subunits.

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Area of Science:

Molecular Biology
Protein Complex Stoichiometry
Yeast Genetics

Background:

The Ski complex in yeast is essential for mRNA degradation, working with the exosome.
Previous understanding suggested a 1:1:1 stoichiometry for Ski2, Ski3, and Ski8 subunits.

Purpose of the Study:

To accurately determine the stoichiometry of the endogenous yeast Ski complex.
To elucidate the subunit arrangement and construct an improved structural model.

Main Methods:

Native mass spectrometry was employed to measure the intact Ski complex.
Tandem mass spectrometry was used to validate subunit interactions and stoichiometry.
Complex disruption and subcomplex mass analysis were performed to probe topology.

Main Results:

Native mass spectrometry demonstrated a hetero-tetrameric stoichiometry: one Ski2, one Ski3, and two Ski8 subunits.
Tandem mass spectrometry confirmed the proposed stoichiometry by analyzing fragment ions.
Analysis of disrupted subcomplexes revealed specific subunit interactions (Ski8-Ski8, Ski2-Ski3, Ski8-Ski2, Ski8-Ski8-Ski2).

Conclusions:

The yeast Ski complex possesses a hetero-tetrameric stoichiometry of Ski2:Ski3:Ski8 in a 1:1:2 ratio.
These findings provide a refined structural model for the Ski complex, crucial for mRNA processing.