From Crystal Structures of RgIA4 in Complex with Ac-AChBP to Molecular Determinants of Its High Potency of α9α10 nAChR.

α9-containing nicotinic acetylcholine receptors (nAChRs) have been shown to play critical roles in neuropathic pain. The α-conotoxin (α-CTx) RgIA and its analog RgIA4 were identified as the most selective inhibitor of α9α10 nAChR. However, the mechanism of their selectivity toward α9α10 nAChR remains elusive. Here, we reported the co-crystal structure of RgIA and RgIA4 in complex with Aplysia californica acetylcholine binding protein (Ac-AChBP) at resolution of 2.6 Å, respectively. Based on the structure of the complexes, together with molecular dynamic simulation (MD-simulation), we suggested the key residues of α9α10 nAChR in determining its high affinity for RgIA/RgIA4. This is the first time the complex between pain-related conotoxins and Ac-AChBP was reported and the complementary side of α9 subunit in binding of the antagonists shown. These results provide realistic template for the design of new therapeutic in neuropathic pain.

Phytoremediation of cadmium contaminated soils by Amaranthus Hypochondriacus L.: The effects of soil properties highlighting cation exchange capacity.

Phytoremediation is a cost-effective method to remedy Cd-contaminated soils. However, it is difficult to predict the performance of a given (hyper)accumulator at different soils due to the divergent plant-soil mutual fitness. Soil properties could be quite influential in determining plant growth and Cd uptake and therefore affect phytoremediation efficiency. To explore the primary soil factors that regulate the efficiency of phytoremediation, a phytoextraction experiment with grain amaranth (Amaranthus Hypochondriacus L.) was conducted in six long-term Cd-contaminated agricultural soils from southern China. The results showed that besides the soil available Cd, the soil cation exchange capacity (CEC) greatly affected plant growth and the amount of total Cd extraction. The deficiency of available Ca and Mg in low CEC soil caused insufficient uptake of Ca and Mg by grain amaranth, which was adverse to plant growth and Cd detoxification. The impaired plant biomass production sharply influenced plant total Cd accumulation, despite the relatively high Cd concentration in plants. While for the grain amaranth grown in soils with higher CEC, the increases in plant Ca and Mg promoted plant photosynthesis and plant tolerance to Cd stress, as indicated by the increase of leaf chlorophyll content and antioxidant enzyme activities, which contributed to the higher plant biomass and phytoremediation efficiency. These findings highlight that maintaining regular plant biomass production is vital to ensure the efficiency of phytoremediation, and low CEC of soil is a substantial barrier that needs to be concerned and further addressed for efficient phytoremediation of Cd-contaminated soils.

High Selectivity of an α-Conotoxin LvIA Analogue for α3ß2 Nicotinic Acetylcholine Receptors Is Mediated by ß2 Functionally Important Residues.

The α3ß2 and α3ß4 nicotinic acetylcholine receptors (nAChRs) are widely expressed in the central and peripheral nervous systems, playing critical roles in various physiological processes and in such pathologies as addiction to nicotine and other drugs of abuse. α-Conotoxin LvIA, which we previously isolated from Conus lividus, modestly discriminates α3ß2 and α3ß4 rat nAChRs exhibiting a ∼17-fold tighter binding to the former. Here, alanine scanning resulted in two more selective analogues [N9A]LvIA and [D11A]LvIA, the former having a >2000-fold higher selectivity for α3ß2. The determined crystal structures of [N9A]LvIA and [D11A]LvIA bound to the acetylcholine-binding protein (AChBP) were followed by homologous modeling of the complexes with the α3ß2 and α3ß4 nAChRs and by receptor mutagenesis, which revealed Phe106, Ser108, Ser113, and Ser168 residues in the ß2 subunit as essential for LvIA binding. These results may be useful for the design of novel compounds of therapeutic potential targeting α3ß2 nAChRs.

Leaf nonstructural carbohydrate concentrations of understory woody species regulated by soil phosphorus availability in a tropical forest.

Leaf soluble sugars and starch are important components of nonstructural carbohydrates (NSCs), which are crucial for plant growth, development, and reproduction. Although there is a large body of research focusing on the regulation of plant NSC (soluble sugars and starch) concentrations, the response of foliar NSC concentrations to continuous nitrogen (N) and phosphorus (P) addition is still unclear, especially in tropical forests. Here, we used a long-term manipulative field experiment to investigate the response of leaf NSC concentrations to continuous N and P addition (3-, 5-, and 8-year fertilization) in a tropical forest in southern China. We found significant species-specific variation in leaf NSC concentrations in this tropical forest. Phosphorus addition dramatically decreased both leaf soluble sugar and starch concentrations, while N addition had no significant effects on leaf soluble sugar and starch concentrations. These results suggest that, in plants growing in P-limiting tropical soil, leaf NSC concentrations are regulated by soil P availability rather than N availability. Moreover, the negative relationships between NSC concentrations and leaf mass per area (LMA) revealed that NSCs could supply excess carbon (C) for leaf expansion under P addition. This was further supported by the increased structural P fraction after P fertilization in our previous study at the same site. We conclude that soil P availability strongly regulates leaf starch and soluble sugar concentrations in the tropical tree species included in this study. The response of leaf NSC concentrations to long-term N and P addition can reflect the close relationships between plant C dynamics and soil nutrient availability in tropical forests. Maintaining relatively higher leaf NSC concentrations in tropical plants can be a potential mechanism for adapting to P-deficient conditions.

The R-loop is a common chromatin feature of the Arabidopsis genome.

R-loops are functional structures in chromatin comprising one single-stranded DNA and a DNA:RNA hybrid. Here, we report ssDRIP-seq, a single-strand DNA ligation-based library preparation technique for genome-wide identification of R-loops. When applied in Arabidopsis, ssDRIP-seq exhibits high efficiency, low bias and strand specificity. We found that Arabidopsis R-loops are enriched by both AT and GC skews, and are formed in the sense and antisense orientations. R-loops are strongly enriched in gene promoters and gene bodies, and are highly associated with noncoding RNA and repetitive genomic regions. Furthermore, R-loops are negatively correlated with CG DNA hypermethylation, and are prevalent in regions with multiple chromatin modifications, showing strong correlations with the activated and repressed gene loci. Our analyses indicate that R-loops are common features in the Arabidopsis genome and suggest that the R-loops play diverse roles in genome organization and gene regulation, thereby providing insights into plant nuclear genome formation and function.