Specific substitution into the anticodon loop of yeast tyrosine transfer RNA

The aminoacylation kinetics of 19 different variants of yeast tRNATyr with nucleotide substitutions in positions 33-35 were determined. Substitution of the conserved uridine-33 does not alter the rate of aminoacylation. However, substitution of the anticodon position 34 or position 35 reduces Km from 2- to 10-fold and Vmax as much as 2-fold, depending on the nucleotide inserted. The ochre and amber suppressor tRNAsTyr both showed about a 7-fold reduction in Vmax/Km. Data from tRNATyr with different modified nucleotides at position 35 suggest that specific hydrogen bonds form between the synthetase and both the N1 and N3 hydrogens of psi-35. The effect of simultaneous substitutions at positions 34 and 35 can be predicted reasonably well by combining the effects of single substitutions. These data suggest that yeast tyrosyl-tRNA synthetase interacts with positions 34 and 35 of the anticodon of tRNATyr and opens the possibility that nonsense suppressor efficiency may be mediated by the level of aminoacylation.     

Physiological and regulatory underpinnings of geographic variation in reptilian cold tolerance across a latitudinal cline

Understanding the mechanisms that produce variation in thermal performance is a key component to investigating climatic effects on evolution and adaptation. However, disentangling the effects of local adaptation and phenotypic plasticity in shaping patterns of geographic variation in natural populations can prove challenging. Additionally, the physiological mechanisms that cause organismal dysfunction at extreme temperatures are still largely under debate. Using the green anole, Anolis carolinensis, we integrate measures of cold tolerance (CT_min), standard metabolic rate, heart size, blood lactate concentration and RNAseq data from liver tissue to investigate geographic variation in cold tolerance and its underlying mechanisms along a latitudinal cline. We found significant effects of thermal acclimation and latitude of origin on variation in cold tolerance. Increased cold tolerance correlates with decreased rates of oxygen consumption and blood lactate concentration (a proxy for oxygen limitation), suggesting elevated performance is associated with improved oxygen economy during cold exposure. Consistent with these results, co‐expression modules associated with blood lactate concentration are enriched for functions associated with blood circulation, coagulation and clotting. Expression of these modules correlates with thermal acclimation and latitude of origin. Our findings support the oxygen and capacity‐limited thermal tolerance hypothesis as a potential contributor to variation in reptilian cold tolerance. Moreover, differences in gene expression suggest regulation of the blood coagulation cascade may play an important role in reptilian cold tolerance and may be the target of natural selection in populations inhabiting colder environments.     

Life cycle assessment capacity roadmap (section 1): decision-making support using LCA

When life cycle assessment (LCA) results do not show a clear and certain environmental preference of one choice over one or several alternatives, current methods are limited in their ability to inform decision-makers. To address this and related cross-cutting issues, a group of LCA practitioners has been working on a roadmap for capacity development in LCA. The roadmap is identifying common needs for development in LCA, which can then be addressed by the broader LCA community. The roadmap document on decision-making support, having undergone a public comment period, outlines the current state as well as needs and milestones to ensure progress continues apace. The roadmap document, available for download, covers five main areas of development: (1) performance measures of confidence, which identify the acceptable uncertainty for study results, while minimizing expenditures; (2) selection of impact categories, an area with multiple existing methods. The roadmap suggests codifying these methods and identifying their suitability to various applications; (3) normalization; while several methods of normalization are in use, the method with the greatest acceptance in the LCA community (i.e., relying on total or per capita regional emissions/extractions) has a number of methodological drawbacks; (4) weighting, which is a form of multi-criteria decision analysis (MCDA). The broader MCDA field can enrich LCA by providing studied methods of assessing trade-offs; and (5) visualization of results. Many other LCA capacity needs would benefit from documentation. These include but are not limited to the following: addressing ill-characterized uncertainty, life cycle inventory data needs, data format needs, and tool capabilities. Other roadmapping groups are forming and are looking for practitioners to support the effort.     

Mutations in DNAJC5, encoding cysteine-string protein alpha, cause autosomal-dominant adult-onset neuronal ceroid lipofuscinosis

Autosomal-dominant adult-onset neuronal ceroid lipofuscinosis (ANCL) is characterized by accumulation of autofluorescent storage material in neural tissues and neurodegeneration and has an age of onset in the third decade of life or later. The genetic and molecular basis of the disease has remained unknown for many years. We carried out linkage mapping, gene-expression analysis, exome sequencing, and candidate-gene sequencing in affected individuals from 20 families and/or individuals with simplex cases; we identified in five individuals one of two disease-causing mutations, c.346_348delCTC and c.344T>G, in DNAJC5 encoding cysteine-string protein alpha (CSPα). These mutations-causing a deletion, p.Leu116del, and an amino acid exchange, p.Leu115Arg, respectively-are located within the cysteine-string domain of the protein and affect both palmitoylation-dependent sorting and the amount of CSPα in neuronal cells. The resulting depletion of functional CSPα might cause in parallel the presynaptic dysfunction and the progressive neurodegeneration observed in affected individuals and lysosomal accumulation of misfolded and proteolysis-resistant proteins in the form of characteristic ceroid deposits in neurons. Our work represents an important step in the genetic dissection of a genetically heterogeneous group of ANCLs. It also confirms a neuroprotective role for CSPα in humans and demonstrates the need for detailed investigation of CSPα in the neuronal ceroid lipofuscinoses and other neurodegenerative diseases presenting with neuronal protein aggregation.     

Hormonal control of morphogenesis in leaf segments ofCentaurium erythraea

Transversally cut leaf segments ofCentaurium erythraea were cultivated on MS medium. Effects of segment polarity, IAA and sucrose concentrations, light and medium volume on morphogenesis were studied. Shoots generally formed at lower (1.3 × 10^-6 mol 1^-1) IAA concentrations than roots and callus (1.1 × 10^-5−3.4 × 10^-5 mol 1^-1). Leaf polarity strongly modified the effect of IAA concentration, shifting organogenesis at the segment base toward decreased IAA concentrations as compared with segment apex. Light, sucrose concentration above 3 % and high medium volume changed IAA dependence of morphogenesis in various ways and generally suppressed segment polarity.