NASP maintains histone H3–H4 homeostasis through two distinct H3 binding modes

Histone chaperones regulate all aspects of histone metabolism. NASP is a major histone chaperone for H3–H4 dimers critical for preventing histone degradation. Here, we identify two distinct histone binding modes of NASP and reveal how they cooperate to ensure histone H3–H4 supply. We determine the structures of a sNASP dimer, a complex of a sNASP dimer with two H3 α3 peptides, and the sNASP–H3–H4–ASF1b co-chaperone complex. This captures distinct functionalities of NASP and identifies two distinct binding modes involving the H3 α3 helix and the H3 αN region, respectively. Functional studies demonstrate the H3 αN-interaction represents the major binding mode of NASP in cells and shielding of the H3 αN region by NASP is essential in maintaining the H3–H4 histone soluble pool. In conclusion, our studies uncover the molecular basis of NASP as a major H3–H4 chaperone in guarding histone homeostasis.     

Adaptive Evolution of cry Genes in Bacillus thuringiensis： Implications for Their Specificity Determination

The cry gene family, produced during the late exponential phase of growth in Bacillus thuringiensis, is a large, still-growing family of homologous genes, in which each gene encodes a protein with strong specific activity against only one or a few insect species. Extensive studies are mostly focusing on the structural and functional relationships of Cry proteins, and have revealed several residues or domains that are important for the target recognition and receptor attachment. In this study, we have employed a maximum likelihood method to detect evidence of adaptive evolution in Cry proteins, and have identified 24 positively selected residues, which are all located in Domain Ⅱ or Ⅲ. Combined with known data from mutagenesis studies, the majority of these residues, at the molecular level, contribute much to the insect specificity determination. We postulate that the potential pressures driving the diversification of Cry proteins may be in an attempt to adapt for the ＂arm race＂ between δ-endotoxins and the targeted insects, or to enlarge their target spectra, hence result in the functional divergence. The sites identified to be under positive selection would provide targets for further structural and functional analyses on Cry proteins.     

Expressing MLH1 in HCT116 cells increases cellular resistance to radiation by activating the PRKAC

Mut L homolog-1 (MLH1) is a key DNA mismatch repair protein which participates in the sensitivity to DNA damaging agents. However, its role in the radiosensitivity of tumor cells is less well characterized. In this study, we investigated the role of MLH1 in cellular responses to ionizing radiation (IR) and explored the signaling molecules involved. The isogenic pair of MLH1 proficient (MLH1⁺) and deficient (MLH1^–) human colorectal cancer HCT116 cells was exposed to IR for 24 h at the dose of 3 cGy. The clonogenic survival was examined by the colony formation assay. Cell cycle distribution was analyzed with flow cytometry. Changes in the protein level of MLH1, DNA damage marker γH2AX, and protein kinase A catalytic subunit (PRKAC), a common target for anti-tumor drugs, were examined with Western blotting. The results showed that the HCT116 (MLH1⁺) cells demonstrated increased radio-resistance with increased S population, decreased G2 population, a low level of γH2AX, a reduced ratio of phosphorylated PRKACαβ to total PRKAC, and an elevated level of total PRKAC and phosphorylated PRKACβII following IR compared with the HCT116 (MLH1^–) cells. Importantly, silencing PRKAC in HCT116 (MLH1⁺) cells increased the cellular radiosensitivity. In conclusion, MLH1 may increase cellular resistance to IR by activating PRKAC. Our finding is the first to demonstrate the important role of PRKAC in MLH1-mediated radiosensitivity, suggesting that PRKAC has potential as a biomarker and a therapeutic target for increasing radio-sensitization.     

Xenophagy of invasive bacteria is differentially activated and modulated via a TLR-TRAF6-Beclin1 axis in echinoderms

In marine environments, organisms are confronted with numerous microbial challenges, although the differential regulation of xenophagy in response to different pathogenic bacterial species remains relatively unknown. Here, we addressed this issue using Apostichopus japonicus as a model. We identified 39 conserved autophagy-related genes by genome-wide screening, which provided a molecular basis for autophagy regulation in sea cucumbers. Furthermore, xenophagy of two Gram-negative bacteria, Vibrio splendidus and Escherichia coli, but not a Gram-positive bacteria, Micrococcus luteus, was observed in different autophagy assays. Surprisingly, a significantly higher autophagy capacity was found in the E. coli–challenged group than in the V. splendidus–challenged group. To confirm these findings, two different lipopolysaccharides, LPS^{V. splendidus} and LPS^E. coli, were isolated; we found that these LPS species differentially activated coelomocyte xenophagy. To explore the molecular mechanism mediating differential levels of xenophagy, we used an siRNA knockdown assay and confirmed that LPS^{V. splendidus}-mediated xenophagy was dependent on an AjTLR3-mediated pathway, whereas LPS^E. coli-mediated xenophagy was dependent on AjToll. Moreover, the activation of different AjTLRs resulted in AjTRAF6 ubiquitination and subsequent activation of K63-linked ubiquitination of AjBeclin1. Inversely, the LPS^{V. splendidus}-induced AjTLR3 pathway simultaneously activated the expression of AjA20, which reduced the extent of K63-linked ubiquitination of AjBeclin1 and impaired the induction of autophagy; however, this finding was no t evident with LPS^E. coli. Our present results provide the first evidence showing that xenophagy could be differentially induced by different bacterial species to yield differential autophagy levels in echinoderms.     

黄曲霉发酵薯干粉水解液生产L－苹果酸

经选育和诱变得到一株产苹果酸的黄曲霉菌ＴＨ５００７,通过发酵条件的优化,以薯干粉的水解液为主要原料,发酵５ｄＬ－苹果酸可达到６％左右。在总酸中苹果酸含量平均达７２％以上,杂有机酸主要是柠檬酸,延胡索酸的含量在０．０２％以下。补糖发酵比分批发酵产酸性能更佳。     

猪生殖—呼吸道综合征病毒（PRRSV）CH—1a株糖蛋白基因的遗传变异分析

新近发现的猪生殖－呼吸道综合征病毒（ＰＲＲＳＶ）是单股ＲＮＡ病毒,属于不久前成立的动脉炎病毒科,为了比较国内分离的ＣＨ－１ａ株与国外毒株的分子遗传学关系,扩增并克隆了ＰＲＲＳＶ ＣＨ－１ａ株糖蛋白基因ＯＲＦ２￣５,测定了其核苷酸序列。用序列分析软件分析了其编码产物的分子量、等电点、疏水性、抗原性和有关位点,并与国外毒株进行了序列比较和系统发育进化分析。结果表明：ＣＨ－１ａ株与ＶＲ－２３３２株尽管密     

北京西山（卧佛寺附近）人工油松林群落学特性及生物量的研究

28年生人工油松林标准地设置在西山海拔300一350米的南坡与北坡山地。试用了多种数学模型预测人工林乔木层的生物量,应用w(树干)=2.372(D2H)0.664, w(枝条)=1.317(D2H)0.992,w(叶子)=1.417(D2H)0.833和w(根系)=1.593(D2H)0.757幂函数方程,分别获得树干、枝条、叶子和根系的生物量。南、北坡油松林乔木层生物量分别为29.13吨／公顷和42.46吨／公顷。用直接收获法测得不同坡向油松林的灌木层和草本层生物量的季节动态,以8月份生物量计,南、北坡灌木层生物量为8.05吨／公顷和5.78吨／公顷,草本层为0.74吨／公顷和0.77吨／公顷。南,北坡油松林的总生物量为38.08吨／公顷和48.68吨／公顷。