In vivo analysis of cohesin architecture using FRET in the budding yeast Saccharomyces cerevisiae

Autor: Frank Uhlmann, Brian E. Snydsman, John Mc Intyre, Trisha N. Davis, Stefan Weitzer, Eric G D Muller
Rok vydání: 2007
Předmět:
Saccharomyces cerevisiae Proteins
Cohesin complex
Chromosomal Proteins
Non-Histone
Recombinant Fusion Proteins
Saccharomyces cerevisiae
chromosome segregation
cohesin
S. cerevisiae
Cell Cycle Proteins
Article
General Biochemistry
Genetics and Molecular Biology
Chromosome segregation
03 medical and health sciences
0302 clinical medicine
Fluorescence Resonance Energy Transfer
Sister chromatids
Molecular Biology
030304 developmental biology
Genetics
0303 health sciences
General Immunology and Microbiology
biology
Cohesin
Kinetochore
General Neuroscience
SMC protein
Nuclear Proteins
biology.organism_classification
Protein Structure
Tertiary
Protein Subunits
Förster resonance energy transfer
Smc proteins
FRET
Biophysics
biological phenomena
cell phenomena
and immunity
030217 neurology & neurosurgery
Zdroj: The EMBO Journal
ISSN: 1460-2075
0261-4189
DOI: 10.1038/sj.emboj.7601793
Popis: Cohesion between sister chromatids in eukaryotes is mediated by the evolutionarily conserved cohesin complex. Cohesin forms a proteinaceous ring, large enough to trap pairs of replicated sister chromatids. The circumference consists of the Smc1 and Smc3 subunits, while Scc1 is thought to close the ring by bridging the Smc (structural maintenance of chromosomes) ATPase head domains. Little is known about two additional subunits, Scc3 and Pds5, and about possible conformational changes of the complex during the cell cycle. We have employed fluorescence resonance energy transfer (FRET) to analyse interactions within the cohesin complex in live budding yeast. These experiments reveal an unexpected geometry of Scc1 at the Smc heads, and suggest that Pds5 plays a role at the Smc hinge on the opposite side of the ring. Key subunit interactions, including close proximity of the two ATPase heads, are constitutive throughout the cell cycle. This depicts cohesin as a stable molecular machine undergoing only transient conformational changes during binding and dissociation from chromosomes. Using FRET, we did not observe interactions between more than one cohesin complex in vivo.
Databáze: OpenAIRE
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