The Mechanism of ATP-Dependent Allosteric Protection of Akt Kinase Phosphorylation

SummaryKinases use ATP to phosphorylate substrates; recent findings underscore the additional regulatory roles of ATP. Here, we propose a mechanism for allosteric regulation of Akt1 kinase phosphorylation by ATP. Our 4.7-μs molecular dynamics simulations of Akt1 and its mutants in the ATP/ADP bound/unbound states revealed that ATP occupancy of the ATP-binding site stabilizes the closed conformation, allosterically protecting pT308 by restraining phosphatase access and key interconnected residues on the ATP→pT308 allosteric pathway. Following ATP→ADP hydrolysis, pT308 is exposed and readily dephosphorylated. Site-directed mutagenesis validated these predictions and indicated that the mutations do not impair PDK1 and PP2A phosphatase recruitment. We further probed the function of residues around pT308 at the atomic level, and predicted and experimentally confirmed that Akt1H194R/R273H double mutant rescues pathology-related Akt1R273H. Analysis of classical Akt homologs suggests that this mechanism can provide a general model of allosteric kinase regulation by ATP; as such, it offers a potential avenue for allosteric drug discovery

Modeling Rett Syndrome Using TALEN-Edited MECP2 Mutant Cynomolgus Monkeys

Gene-editing technologies have made it feasible to create nonhuman primate models for human genetic disorders. Here, we report detailed genotypes and phenotypes of TALEN-edited MECP2 mutant cynomolgus monkeys serving as a model for a neurodevelopmental disorder, Rett syndrome (RTT), which is caused by loss-of-function mutations in the human MECP2 gene. Male mutant monkeys were embryonic lethal, reiterating that RTT is a disease of females. Through a battery of behavioral analyses, including primate-unique eye-tracking tests, in combination with brain imaging via MRI, we found a series of physiological, behavioral, and structural abnormalities resembling clinical manifestations of RTT. Moreover, blood transcriptome profiling revealed that mutant monkeys resembled RTT patients in immune gene dysregulation. Taken together, the stark similarity in phenotype and/or endophenotype between monkeys and patients suggested that gene-edited RTT founder monkeys would be of value for disease mechanistic studies as well as development of potential therapeutic interventions for RTT

AMS measurement of 53Mn and its initial application at CIAE

The determination of cosmogenic 53Mn in terrestrial archives has important applications, such as burial ages, exposure age and erosion rates. Accelerator mass spectrometry (AMS) is the most sensitive technique to detect minute amounts of 53Mn. 53Mn measurements were developed at the China Institute of Atomic nergy (CIAE) using the DE-Q3D equipped AMS system. This approach was recently optimized with the goal to reach the sensitivity required for AMS measurements of 53Mn in deep-sea ferromanganese crust (DSFC) samples. Based on these improvements of sample preparation, current beam transmission and so on, 53Mn in two samples of DSFC was measured by AMS. The ratios of 53Mn/Mn corresponding to an age of 3.77 ± 0.42 and 13.73 ± 2.74 Ma by 129I dating method are (5.01 ± 2.15) 10 13 and (1.90 ± 0.96) 10 13. The ratios are close to the experimental reference values, deduced from the previous research. The experimental progress, performances and results are presented in this contribution.This work was mainly supported by the National Natural Science Foundations of China (NSFC), under Grant No. 11075221, and a partly supported by the National Natural Science Foundation of China under Grant Nos. 10705054, 41073044 and 11265005

Big-five personality factors affecting pedestrian crossing behaviors

Human personality is an important component of psychological factors affecting pedestrian crossing. This paper reports a questionnaire survey on the effects of pedestrian personalities (including neuroticism, extraversion, openness, agreeableness and conscientiousness) on pedestrian violation in China. 675 feedbacks were obtained, of which 535 samples were valid for analysis. The results of the hierarchical regression analysis showed that educational level had significant effect on violation; agreeableness had significant effect on violation, conditional compliance and unconditional compliance; consciousness had significant effect on violation and conditional compliance; extraversion had significant effect on unconditional compliance; neuroticism had significant effect on violation; educational level had significant effect on violation. The results implied that psychological measures played a very important role in pedestrian safety