N6-methyladenosine in nuclear RNA is a major substrate of the obesity-associated FTO

We report here that fat mass and obesity-associated protein (FTO) has efficient oxidative demethylation activity targeting the abundant N6-methyladenosine (m(6)A) residues in RNA in vitro. FTO knockdown with siRNA led to increased amounts of m(6)A in mRNA, whereas overexpression of FTO resulted in decreased amounts of m(6)A in human cells. We further show the partial colocalization of FTO with nuclear speckles, which supports the notion that m(6)A in nuclear RNA is a major physiological substrate of FTO.     

Potassium Bisperoxo(1,10-phenanthroline)oxovanadate (bpV(phen)) Induces Apoptosis and Pyroptosis and Disrupts the P62-HDAC6 Protein Interaction to Suppress the Acetylated Microtubule-dependent Degradation of Autophagosomes

Autophagy is a cellular process that controls and executes the turnover of dysfunctional organelles and misfolded or abnormally aggregated proteins. Phosphatase and tensin homologue deleted on chromosome 10 (PTEN) activates the initiation of autophagy. Autophagosomes migrate along acetylated microtubules to fuse with lysosomes to execute the degradation of the engulfed substrates that usually bind with sequestosome 1 (SQSTM1, p62). Microtubule-associated protein 1 light chain 3 (LC3) traces the autophagy process by converting from the LC3-I to the LC3-II isoform and serves as a major marker of autophagy flux. Potassium bisperoxo(1,10-phenanthroline)oxovanadate (bpV(phen)) is an insulin mimic and a PTEN inhibitor and has the potential to treat different diseases. Here we show that bpV(phen) enhances the ubiquitination of p62, reduces the stability of p62, disrupts the interaction between p62 and histone deacetylase 6 (HDAC6), activates the deacetylase activity of HDAC6 on α-tubulin, and impairs stable acetylated microtubules. Microtubular destabilization leads to the blockade of autophagosome-lysosome fusion and accumulation of autophagosomes. Autophagy defects lead to oxidative stress and lysosomal rupture, which trigger different types of cell death, including apoptosis and pyroptosis. The consistent results from multiple systems, including mouse and different types of mammalian cells, are different from the predicted function of bpV(phen) as a PTEN inhibitor to activate autophagy flux. In addition, levels of p62 are reduced but not elevated when autophagosomal degradation is blocked, revealing a novel function of p62 in autophagy regulation. Therefore, it is necessary to pay attention to the roles of bpV(phen) in autophagy, apoptosis, and pyroptosis when it is developed as a drug.     

成土时间对黄河三角洲植物及微生物演替的影响

受时空因素的限制,研究土壤、微生物和植物的演替过程通常采用以空间换时间的时空替代法,关于成土时间的驱动力研究较少。选取黄河三角洲4块代表性冲积区域（形成年代分别为1904-1929年,1929-1934年,1964-1976年和1976年至今）,由沿海到农田（1-40 km）设计采样路线,通过多样性调查、土壤采样、MiSeq测序等方法对陆地生态系统的植物、细菌和真菌多样性以及微生物功能组成进行了监测。结果表明,随成土时间的增加,植物物种丰富度和生物量显著提高（P<0.05）,低盐度指示物种白茅的分布区域向海岸推进;细菌群落酸杆菌门（Acidobacteriota）等3个门类占比显著增加,拟杆菌门（Bacteroidota）等12个门类占比显著减少（P<0.05）,涉及基因信息处理、核苷酸代谢等基础生命活动和光合作用的功能通路在年代最晚的扇区富集,而涉及细胞老化调控、外源物质生物降解与代谢、芳香化合物降解等功能的通路在年代最早的扇区富集（LDA score>2,P<0.05）;真菌群落涉及病理营养的捕虫霉门（Zoopagomycota）等类群相对丰度的显著增加（P<0.05）。耐盐菌、自养菌、反硝化菌、反硫化菌等功能群在成土较晚区域的丰富度较高,有促进成土初期土壤碳氮积累、土壤养分循环和有效性的提升的功能潜力,进而影响植物演替。成土时间还驱动适应高营养环境的类群对先锋类群比值的增大,促进微生物生态位的分化。研究表明成土时间是对黄河三角洲植物和微生物群落组成变化的重要而显著的驱动力,为提高评估滨海生态系统植物和微生物演替规律的准确性提供了科学依据。     

Complete Genome Sequence of a Novel Variant Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Strain: Evidence for Recombination between Vaccine and Wild-Type PRRSV Strains

Porcine reproductive and respiratory syndrome virus (PRRSV) is the etiologic agent of porcine reproductive and respiratory syndrome (PRRS), which can evolve continuously by random mutation or intragenic recombination. Here we report the complete genomic sequence of a PRRSV variant with nucleotide acid deletions and insertions in the nonstructural protein 2 (nsp2) gene and a possible recombination event between a modified live virus (MLV) vaccine strain and a prototype Chinese field strain.     

Metabolic Interactions between the Lands Cycle and the Kennedy Pathway of Glycerolipid Synthesis in Arabidopsis Developing Seeds

It has been widely accepted that the primary function of the Lands cycle is to provide a route for acyl remodeling to modify fatty acid (FA) composition of phospholipids derived from the Kennedy pathway. Lysophosphatidylcholine acyltransferase (LPCAT) is an evolutionarily conserved key enzyme in the Lands cycle. In this study, we provide direct evidence that the Arabidopsis thaliana LPCATs, LPCAT1 and LPCAT2, participate in the Lands cycle in developing seeds. In spite of a substantially reduced initial rate of nascent FA incorporation into phosphatidylcholine (PC), the PC level in the double mutant lpcat1 lpcat2-2 remained unchanged. LPCAT deficiency triggered a compensatory response of de novo PC synthesis and a concomitant acceleration of PC turnover that were attributable at least in part to PC deacylation. Acyl-CoA profile analysis revealed complicated metabolic alterations rather than merely reduced acyl group shuffling from PC in the mutant. Shifts in FA stereo-specific distribution in triacylglycerol of the mutant seed suggested a preferential retention of saturated acyl chains at the stereospecific numbering (sn)-1 position from PC and likely a channeling of lysophosphatidic acid, derived from PC, into the Kennedy pathway. Our study thus illustrates an intricate relationship between the Lands cycle and the Kennedy pathway.     

Probing N6-methyladenosine RNA modification status at single nucleotide resolution in mRNA and long noncoding RNA

N⁶-methyladenosine (m⁶A) is the most abundant modification in mammalian mRNA and long noncoding RNA (lncRNA). Recent discoveries of two m⁶A demethylases and cell-type and cell-state-dependent m⁶A patterns indicate that m⁶A modifications are highly dynamic and likely play important biological roles for RNA akin to DNA methylation or histone modification. Proposed functions for m⁶A modification include mRNA splicing, export, stability, and immune tolerance; but m⁶A studies have been hindered by the lack of methods for its identification at single nucleotide resolution. Here, we develop a method that accurately determines m⁶A status at any site in mRNA/lncRNA, termed site-specific cleavage and radioactive-labeling followed by ligation-assisted extraction and thin-layer chromatography (SCARLET). The method determines the precise location of the m⁶A residue and its modification fraction, which are crucial parameters in probing the cellular dynamics of m⁶A modification. We applied the method to determine the m⁶A status at several sites in two human lncRNAs and three human mRNAs and found that m⁶A fraction varies between 6% and 80% among these sites. We also found that many m⁶A candidate sites in these RNAs are however not modified. The precise determination of m⁶A status in a long noncoding RNA also enables the identification of an m⁶A-containing RNA structural motif.     

Curcumin prevents obesity by targeting TRAF4‐induced ubiquitylation in m6A‐dependent manner

Obesity has become a major health problem that has rapidly prevailed over the past several decades worldwide. Curcumin, a natural polyphenolic compound present in turmeric, has been shown to have a protective effect on against obesity and metabolic diseases. However, its underlying mechanism remains largely unknown. Here, we show that the administration of curcumin significantly prevents HFD‐induced obesity and decreases the fat mass of the subcutaneous inguinal WAT (iWAT) and visceral epididymal WAT (eWAT) in mice. Mechanistically, curcumin inhibits adipogenesis by reducing the expression of AlkB homolog 5 (ALKHB5), an m⁶A demethylase, which leads to higher m⁶A‐modified TNF receptor‐associated factor 4 (TRAF4) mRNA. TRAF4 mRNA with higher m⁶A level is recognized and bound by YTHDF1, leading to enhanced translation of TRAF4. TRAF4, acting as an E3 RING ubiquitin ligase, promotes degradation of adipocyte differentiation regulator PPARγ by a ubiquitin–proteasome pathway thereby inhibiting adipogenesis. Thus, m⁶A‐dependent TRAF4 expression upregulation by ALKBH5 and YTHDF1 contributes to curcumin‐induced obesity prevention. Our findings provide mechanistic insights into how m⁶A is involved in the anti‐obesity effect of curcumin.