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Although mistranslation is commonly believed to be deleterious, recent evidence indicates that mistranslation can be actively regulated and be beneficial in stress response. Methionine mistranslation in mammalian cells is regulated by reactive oxygen species where cells deliberately alter the proteome through incorporating Met at non-Met positions to enhance oxidative stress response. However, it was not known whether specific, mistranslated mutant proteins have distinct activities from the wild-type protein whose sequence is restrained by the genetic code. Here, we show that Met mistranslation with and without Ca(2+) overload generates specific mutant Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) proteins substituting non-Met with Met at multiple locations. Compared to the genetically encoded wild-type CaMKII, specific mutant CaMKIIs can have distinct activation profiles, intracellular localization and enhanced phenotypes. Our results demonstrate that Met-mistranslation, or "Met-scan" can indeed generate mutant proteins in cells that expand the activity profile of the wild-type protein, and provide a molecular mechanism for the role of regulated mistranslation.

There is increasing evidence of environmental change impacts on ecosystem processes and services, yet poor understanding of the relative contributions of land use and climate change to ecosystem services variations. Based on detailed meteorological, hydrological records and satellite data over the Yellow River Source Area (YRSA) in Tibetan Plateau from 1980s to 2008, together with a water-yield module of Integrated Valuation of Ecosystem Services and Tradeoffs (InVEST) model and also a Residual Trends (RESTREND) method, we assessed the water supply variations in YRSA during the past three decades and disentangled the relative contributions of land use and climate change. Results show that water supply significantly decreased from 1980 to 2005 and then increased from 2005 to 2008. The quantity slightly decreased from 283.01 mm in 1980 to 276.95 mm in 1995, 270.12 mm in 2000 and 267.97 mm in 2005, and it then rebounded slightly to 275.26 mm in 2008. The water supply variation ranged from 283.01 mm to 267.97 mm. Climate change contributed dominantly to water supply decrease from 1980 to 1995, which accounts for approximately 64% of the decrease. During 1995 to 2000, land use contributed more and about 58% to the water supply decrease as the intense human activities. From 2000 to 2005, climate change became a positive contribution to the water supply as the increased precipitation, but the land use still contributed negatively. From 2005 to 2008, both climate and land use have positive impacts, but land use contributed about 61% to the water supply increase. The implementation of the Three Rivers Source Area Ecological Protection Project has greatly improved the vegetation coverage conditions and the water retention ability during this period. We recommend that the implementation of ecological projects, grazing policies and artificial improvement of degraded grassland would help to conserve the water retention ability and increase water supply.

Objective The objective of this biomechanical study was to evaluate the stability provided by a newly developed shape memory alloy hook (SMAH) in a cadaveric transforaminal lumbar interbody fusion (TLIF) model. Methods Six human cadaveric spines (L1-S2) were tested in an in vitro flexibility experiment by applying pure moments of ±8 Nm in flexion/extension, left/right lateral bending, and left/right axial rotation. After intact testing, a TLIF was performed at L4-5. Each specimen was tested for the following constructs: unilateral SMAH (USMAH); bilateral SMAH (BSMAH); unilateral pedicle screws and rods (UPS); and bilateral pedicle screws and rods (BPS). The L3–L4, L4–L5, and L5-S1 range of motion (ROM) were recorded by a Motion Analysis System. Results Compared to the other constructs, the BPS provided the most stability. The UPS significantly reduced the ROM in extension/flexion and lateral bending; the BSMAH significantly reduced the ROM in extension/flexion, lateral bending, and axial rotation; and the USMAH significantly reduced the ROM in flexion and left lateral bending compared with the intact spine (p0.05). Stability provided by the USMAH compared with the UPS was not significantly different. ROMs of adjacent segments increased in all fixed constructs (p>0.05). Conclusions Bilateral SMAH fixation can achieve immediate stability after L4–5 TLIF in vitro. Further studies are required to determine whether the SMAH can achieve fusion in vivo and alleviate adjacent segment degeneration.

Epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I) are critical regulators of cell differentiation, survival, proliferation, and migration in cancers. This study found that ARNO (cytohesin-2), an activator of the EGF and IGF-I pathways, was more highly expressed in colorectal cancer tissue than in benign adjacent colorectal tissue. When ARNO-siRNA or the chemical inhibitor SecinH3 blocked ARNO, the downstream of the EGF and IGF-I pathways decreased in colorectal cell lines HT29 and HCT116. This blocking also weakened cell proliferation, invasion, and migration in vitro. Furthermore, EGF receptor (EGFR)-dependent colorectal tumor xenografts in nude mouse exerted anti-proliferative and growth suppression effects by injecting secineH3. These data suggested that inhibiting cytohesins or ARNO as cytoplasmic activators of EGFR and IGF-I in colorectal cancer resulted in anti-proliferation, reduced invasion, decreased migration, and suppressed growth in vivo and in vitro. Therefore, cytohesins or ARNO may be a potential therapy target for some colorectal cancer.

Stress-induced changes of gene expression are crucial for survival of eukaryotic cells. Regulation at the level of translation provides the necessary plasticity for immediate changes of cellular activities and protein levels. In this study, we demonstrate that exposure to oxidative stress results in a quick repression of translation by deactivation of the aminoacyl-ends of all transfer-RNA (tRNA). An oxidative-stress activated nuclease, angiogenin, cleaves first within the conserved single-stranded 3′-CCA termini of all tRNAs, thereby blocking their use in translation. This CCA deactivation is reversible and quickly repairable by the CCA-adding enzyme [ATP(CTP):tRNA nucleotidyltransferase]. Through this mechanism the eukaryotic cell dynamically represses and reactivates translation at low metabolic costs., Author Summary Adequate reprogramming of metabolic activities by environmental stress or suboptimal growth conditions is crucial for cell survival. Cells employ a remarkable diversity of processes to maintain its homeostasis at all levels of gene expression, including chromatin remodeling, mRNA expression and degradation, translation and protein degradation. Each of these processes shapes cell response at different time scales. In this study, we analyzed the cellular response to oxidative stress at the level of translation. Translation, as one of the most downstream processes in gene expression, provides the necessary plasticity for immediate changes of cellular activities. Using high-sensitive approaches to probe the structural integrity of cellular tRNAs, we show that upon exposure to oxidative stress tRNAs are rapidly deactivated by a cleavage within their ubiquitous, single-stranded 3′-CCA termini by oxidative stress-activated nuclease, angiogenin. The CCA-ends deactivation is reversible and quickly repairable by a ubiquitous enzyme, CCA-adding enzyme, whose natural function is to attach post-transcriptionally the CCA overhang to the 3′-termini of all tRNAs in an mRNA template-independent manner. We propose that this is a mechanism to dynamically repress and reactivate translation at low metabolic costs.