To test whether increased α3 connexin proteins expressing from knock-in α8(Kiα3/Kiα3) alleles can prevent a severe nuclear cataract caused by the γB-crystallin S11R mutant proteins expressing from γB S11R/S11R) alleles, we generated compound mutant γB S11R/S11R) α8(Kiα3/Kiα3) mice.
This result is extremely interesting as it suggests either that the lentiviral particles diffused throughout the brain or that the mutant proteins expressed following virion integration, were propagated from cell to cell, possibly through exosomes [37].
All of the mutant proteins expressed at levels similar to wild type and accumulated as native and soluble proteins.
All three mutant proteins expressed with Upf2p failed to promote growth on selective media indicating a significant decrease in the strength of the interaction (Table 2, (7)).
The corresponding mutations were introduced into the respective cDNAs by site directed mutagenesis and the mutant proteins expressed in HaCaT cells.
As a consequence, the RAS mutant proteins expressed in cancer cells remain in the active GTP-bound form and both cell growth and apoptosis regulatory pathways are scrambled.
They found that resveratrol reduced significantly the extent of neuronal cell death and neurotoxicity of the mutant proteins expressed in these two models (p25, a toxic coactivator of CDK5, and a mutant form of superoxide dismutase 1, respectively).
It has been shown previously that some CBS mutant proteins expressed in bacteria misassemble and aggregate [ de Franchis et al., 1999; Gaustadnes et al., 2002; Janosik et al., 2001b; Singh et al., 2007; Yamanishi et al., 2006].
The functional significance of the 12 variant sequences has been investigated in vitro using mutant proteins expressed from ZIC3 full-length cDNAs containing each relevant mutation (Ware et al., 2004; Chhin et al., 2007).
A E357K mutation was made by site directed mutagenesis and the mutant protein expressed and purified to >95% purity as assessed by SDS-PAGE analysis (data not shown).
