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Individual SCF Pop1pand SCF Pop2pcomplexes bearing ubiquitin ligase activity
In the wild type lenses, typical cross-sectional patterns of fiber cells organized in radial columns were observed (Figure 3K,3N,3Pand 3S).
While these data support the existence of distinct SCF Pop1pand SCF Pop2pcomplexes, individual binding of Pop1p and Pop2p to SCF core components in the absence of their heterodimerizing F-box protein partners does not rule out the possibility that these complexes represent inactive intermediates formed during the normal assembly of functional SCF Pop1p-Pop2pcomplexes.
To exclude this possibility, we asked whether distinct SCF Pop1pand SCF Pop2pcomplexes bear ubiquitin ligase activity in vitro . To this end, we performed in vitro ubiquitylation assays.
These data indicate that Pop1p and Pop2p assume their subcellular localization pattern independent of each other, indicating the possibility of distinct nuclear and cytoplasmic homooligomeric SCF Pop1pand SCF Pop2pcomplexes.