Note for: A comprehensive framework of E2-RING E3 interactions of the human ubiquitin-proteasome system
Skimming: A comprehensive framework of E2-RING E3 interactions of the human ubiquitin-proteasome system
(doi: 10.1038/msb.2009.55)
Using yeast two-hybrid (Y2H) screening for the interaction partner between E2-E3 and vice versa
Reporter system:
LacZ
LEU2
Human cDNA-->produce only the domain (partial)
E2 -- UBC-fold (highly conserved)
E3 -- RING-finger catalytic domain (interact with E2 through the UBC fold)
E2 interacts more than one E3 and vice versa
size of circles reflects the numbers of interaction partners of either E2 (red) or E3 (blue).
Environment to examine the interaction
in yeast without any stimulant
Platform of yeast two-hybrid screening
Each domain connected to reporter protein at the C-terminal//each transform to yeast (haploid stage)//mating between two different sexual strain. If there is the interaction between E2-E3-->yeast can grow on the selective media.
(doi: 10.1038/msb.2009.55)
Using yeast two-hybrid (Y2H) screening for the interaction partner between E2-E3 and vice versa
Reporter system:
LacZ
LEU2
Human cDNA-->produce only the domain (partial)
E2 -- UBC-fold (highly conserved)
E3 -- RING-finger catalytic domain (interact with E2 through the UBC fold)
E2 interacts more than one E3 and vice versa
size of circles reflects the numbers of interaction partners of either E2 (red) or E3 (blue).
Environment to examine the interaction
in yeast without any stimulant
Platform of yeast two-hybrid screening
Each domain connected to reporter protein at the C-terminal//each transform to yeast (haploid stage)//mating between two different sexual strain. If there is the interaction between E2-E3-->yeast can grow on the selective media.
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