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Lupski's three sick siblings also had both of these mutations, whereas his four healthy siblings and parents (who do not have the disease) carried only one mutated gene.
Using biochemical assays we demonstrated that both of these mutations led to significantly reduced Sin1 phosphorylation at the T86 site, as these mutations impair the canonical AGC kinase consensus recognition motif "RxRxxpS/pT" (Alessi et al., 1996; Manning and Cantley, 2007).
Both of these mutations were also detected in the corresponding tumor (Fig. 2E).
As shown in Figure 2D, both of these mutations essentially abolished modification of this construct.
Considering that both of these mutations can alter the structure, this finding may indicate that the sites co-evolve to maintain a specific protein conformation.
The Iowa and Flemish mutations did not appear to result in different morphologies of fibrils compared to Wild Type; however, the extent of fibril formation appeared to be reduced in the presence of the bilayer by both of these mutations.
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We therefore wondered whether the loss of the 85 kb fragment and the truncation of cre1 were the result of one or both of these mutation steps.
The rate of conversion of DNA adducts into mutations, estimated using the ratio of mutant frequency to adduct frequency, and denoted as mutagenic efficiency, was calculated for both of these mutation targets in order to compare susceptibility of the two loci.
Furthermore, database searches reveal that truncating mutations, in reportedly phenotypically normal individuals, can occur both 5' and 3' of these mutations, just as we now report for ASXL3 (see Additional file 7, Figure S5).
Future research will be designed to definitively correlate one or both of these genetic mutations to the enhanced xylose usage phenotype seen in the newly generated strains.
These enzymatic data indicate that both of these histidine mutations when introduced into human PLCζ dramatically diminish their PIP2 hydrolytic activity, thus directly explaining why the cRNA microinjection of these PLCζ mutants into unfertilized mouse eggs fails to induce normal egg activation.
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