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dPANK/Fbl Drosophila mutants possess a neurodegenerative phenotype and a greatly reduced life span.
However, only mpk4 mutants possess a similar phenotype to mekk1 and have similar changes in gene expression.
Although, both the mutants possess a similar secondary structural content, the stability of ATPase domain of the R126W variant is significantly altered.
Consistent with this hypothesis, many human tumour-associated p53 mutants possess a number of properties absent from the wild-type protein [ 3].
Prior studies have suggested that daf-2 (e1370 ) mutants possess a metabolic profile that is different from that of wild type (Murphy et al. 2003; Houthoofd et al. 2005; Mendenhall et al. 2006).
In support of the ATPase requirement are mutational studies illustrating that walker-type ATP binding site mutants possess a dominant-negative phenotype (Bamming and Horvath 2009; Yoneyama et al. 2004).
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In order to verify none of the spontaneous resistance mutants possessed a general growth defect, the growth rate was determined at 30°C in a gyrator shaker in LB [24], [58].
After six rounds of combined mutagenesis and DNA shuffling, seven mutants possessing a total of six point mutations were identified.
As a comparison, among the previously studied S/T protein kinase knockouts in N. crassa (Park et al. 2011b), 57% of the mutants possessed a defect in at least one of the growth/developmental stages, whereas 45% had overlapping defects in all three.
Further investigation showed that the P474N mutant possessed a higher (13.3%, p < 0.05) specific activity.
The c68 (docs1 1) mutant possessed a nonsense mutation in the C-terminal part of the DOCS1 kinase domain.
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