Multicomponent Yeast Two-Hybrid System: Applications to Study Protein-Protein Interactions in SMC Complexes
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
- Keywords
- KITE proteins, Kleisin bridge, Multicomponent yeast two-hybrid (Y2H) system, Non-SMC Element (Nse) subunits, Protein–protein interactions, SMC5/6, Structural maintenance of chromosomes (SMC) complexes,
- MeSH
- Chromosomes, Fungal physiology MeSH
- Protein Interaction Maps physiology MeSH
- Multiprotein Complexes metabolism MeSH
- Protein Subunits metabolism MeSH
- Cell Cycle Proteins metabolism MeSH
- Saccharomyces cerevisiae Proteins metabolism MeSH
- Saccharomyces cerevisiae metabolism MeSH
- Two-Hybrid System Techniques MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Multiprotein Complexes MeSH
- Protein Subunits MeSH
- Cell Cycle Proteins MeSH
- Saccharomyces cerevisiae Proteins MeSH
Analysis of protein-protein interactions (PPI) is key for the understanding of most protein assemblies including structural maintenance of chromosomes (SMC) complexes. SMC complexes are composed of SMC proteins, kleisin, and kleisin-interacting subunits. These subunits interact in specific ways to constitute and regulate the closed structure of the complexes. Specifically, kleisin molecules bridge the SMC dimers and the kleisin-interacting subunits modulate stability of the bridge. Here we describe a multicomponent version of a yeast two-hybrid (Y2H) method and its application for analysis of the bridging role of the Nse4 kleisin in the SMC5/6 complex. Using this technique, we also show a stabilizing effect of KITE (kleisin-interacting tandem winged-helix element) proteins on SMC5/6.
Genome Damage and Stability Centre University of Sussex Brighton UK
National Centre for Biomolecular Research Faculty of Science Masaryk University Brno Czech Republic
References provided by Crossref.org
Characterisation of the Arabidopsis thaliana telomerase TERT-TR complex
The SAGA histone acetyltransferase module targets SMC5/6 to specific genes
A role of the Nse4 kleisin and Nse1/Nse3 KITE subunits in the ATPase cycle of SMC5/6
