Mechanistic insights into tRNA cleavage by a contact-dependent growth inhibitor protein and translation factors

Contact-dependent growth inhibition is a mechanism of interbacterial competition mediated by delivery of the C-terminal toxin domain of CdiA protein (CdiA–CT) into neighboring bacteria. The CdiA–CT of enterohemorrhagic Escherichia coli EC869 (CdiA–CT^EC869) cleaves the 3′-acceptor regions of specific tRNAs in a reaction that requires the translation factors Tu/Ts and GTP. Here, we show that CdiA–CT^EC869 has an intrinsic ability to recognize a specific sequence in substrate tRNAs, and Tu:Ts complex promotes tRNA cleavage by CdiA–CT^EC869. Uncharged and aminoacylated tRNAs (aa-tRNAs) were cleaved by CdiA–CT^EC869 to the same extent in the presence of Tu/Ts, and the CdiA–CT^EC869:Tu:Ts:tRNA(aa-tRNA) complex formed in the presence of GTP. CdiA–CT^EC869 interacts with domain II of Tu, thereby preventing the 3′-moiety of tRNA to bind to Tu as in canonical Tu:GTP:aa-tRNA complexes. Superimposition of the Tu:GTP:aa-tRNA structure onto the CdiA–CT^EC869:Tu structure suggests that the 3′-portion of tRNA relocates into the CdiA–CT^EC869 active site, located on the opposite side to the CdiA–CT^EC869 :Tu interface, for tRNA cleavage. Thus, CdiA–CT^EC869 is recruited to Tu:GTP:Ts, and CdiA–CT:Tu:GTP:Ts recognizes substrate tRNAs and cleaves them. Tu:GTP:Ts serves as a reaction scaffold that increases the affinity of CdiA–CT^EC869 for substrate tRNAs and induces a structural change of tRNAs for efficient cleavage by CdiA–CT^EC869.     

Nitidine chloride inhibited the expression of S phase kinase-associated protein 2 in ovarian cancer cells

Nitidine chloride (NC) has been reported to exert its anti-tumor activity in various types of human cancers. However, the molecular mechanism of NC-mediated tumor suppressive function is largely unclear. In the current study, we used several approaches such as MTT, FACS, RT-PCR, Western blotting analysis, invasion assay, transfection, to explore the molecular basis of NC-triggered anti-cancer activity. We found that NC inhibited cell growth, induced cell apoptosis, caused cell cycle arrest in ovarian cancer cells. Emerging evidence has demonstrated that Skp2 plays an important oncogenic role in ovarian cancer. Therefore, we also explored whether NC exerts its biologic function via downregulation of Skp2 in ovarian cancer cells. We observed that NC significantly inhibited the expression of Skp2 in ovarian cancer cells. Notably, overexpression of Skp2 abrogated the anti-cancer activity induced by NC in ovarian cancer cells. Consistently, downregulation of Skp2 expression enhanced the sensitivity of ovarian cancer cells to NC treatment. Thus, inactivation of Skp2 by NC could be a novel strategy for the treatment of human ovarian cancer.     

利用癌功能类从癌基因组突变谱中识别癌基因

从大规模癌样本基因突变扫查数据中识别癌基因具有重要的意义. 一些重要功能的改变对于癌的发生发展是必需的, 因此将它们定义为癌功能类, 并从GO(Gene Ontology)中选择一组显著富集已知癌基因的细致功能类来代表它们. 为了评价以癌相关功能类作为特征识别癌基因的效果, 将已知的蛋白激酶癌基因定义为阳性金标准, 而将其他的蛋白激酶基因定义为阴性金标准. 结果表明, 与利用选择压力作为特征的方法比较, 利用癌相关功能类作为特征的方法可以更有效地识别癌基因. 进一步结合癌相关功能类与基因非同义突变个数可以产生更可靠的预测结果. 最后, 将46个注释到癌相关功能类并且其非同义突变个数至少为3的蛋白激酶基因预测为癌基因, 预测精确率达到0.42.     

栗山天牛天敌花绒寄甲在栎林中的种群保持机制

花绒寄甲(Dastarcus helophoroides)(鞘翅目:寄甲科Bothrideridae)是寄生栗山天牛(Massicus raddei)中老龄幼虫和蛹的重要天敌,但其寄主栗山天牛世代周期长(3年1代)、发育比较整齐,不利于寄生性天敌种群数量的稳定.为了解利用花绒寄甲防治栗山天牛后,其种群能否在栎树林间长期保持较高的种群数量,达到持续控制栗山天牛的防治效果,调查研究了花绒寄甲在栎树林间的转主寄主和种群保持机制.结果表明,在东北辽东栎树干和树枝上除了栗山天牛外,还有其他8种天牛危害:双簇天牛(Moechotypa diphysis)、四点象天牛(Mesosa myops)、中华薄翅锯天牛(Megopis sinica)、锯天牛(Prionus insularis)、双带粒翅天牛(Lamiomimus gottschei)、八字绿虎天牛(Chlorophorus tohokensis)、日本绿虎天牛(C.japonicus)和拟蜡天牛(Stenygrinumquadrinotatum).其中以栗山天牛、双簇天牛、四点象天牛和拟蜡天牛数量较多,而花绒寄甲在辽东栎树干上的垂直分布与栗山天牛、双簇天牛和四点象天牛的垂直分布重叠较多.室内研究表明,花绒寄甲对四点象天牛老熟幼虫的寄生率达到26.67％,对蛹的寄生率达到了43.33％;对双簇天牛老熟幼虫的寄生率达到20.00％,对蛹的寄生率为6.67％.对双簇天牛和四点象天牛在林间的生活史调查和研究发现,花绒寄甲可寄生的这两种天牛的中老龄幼虫和蛹,在花绒寄甲不适宜寄生的栗山天牛幼龄幼虫期大量存在,表明双簇天牛和四点象天牛是花绒寄甲在栎树林中的主要转主寄主.由于这些转主寄主的存在,花绒寄甲在不利于其寄生的栗山天牛卵期、幼龄幼虫期可转移寄生这些寄主,从而在栗山天牛危害的栎树林间保持了较高的种群数量,达到对栗山天牛长期而有效的持续控制效果.     

The structure of the antimicrobial peptide Ac-RRWWRF-NH2 bound to micelles and its interactions with phospholipid bilayers.

The hexapeptide Ac-RRWWRF-NH2 has earlier been identified as a potent antimicrobial peptide by screening synthetic combinatorial hexapeptide libraries. In this study, it was found that this peptide had a large influence on the thermotropic phase behavior of model membranes containing the negatively charged headgroup phosphatidylglycerol, a major component of bacterial membranes. In contrast, differential scanning calorimetry showed that it had little effect on model membranes containing the zwitterionic phosphatidylcholine headgroup, the main component of erythrocyte membranes. This behavior is consistent with its biological activity and with its affinity to these membranes as determined by titration calorimetry, implying that peptide-lipid interactions play an important role in this process. The structure of this peptide bound to membrane-mimetic sodium dodecyl sulfate (SDS) and dodecylphosphocholine micelles has been determined using conventional two-dimensional nuclear magnetic resonance methods. It forms a marked amphipathic structure in SDS with its hydrophobic residues on one side of the structure and with the positively charged residues on the other side. This amphipathic structure may allow this peptide to penetrate deeper into the interfacial region of negatively charged membranes, leading to local membrane destabilization. Knowledge about the importance of electrostatic interactions of Arg and the role of Trp residues as a membrane interface anchor will provide insight into the future design of potent antimicrobial peptidomimetics.     

SCREENING OF IRON- AND ZINC-ENRICHED YEAST STRAIN AND OPTIMIZATION OF CULTIVATION CONDITIONS

Saccharomyces cerevisiae LN-17 was selected from 26 kinds of primary yeast strains that belong to different genera and species. The iron- and zinc-enriched capability of strain LN-17 was higher than the others. The highest iron and zinc contents of the strain were obtained when the strain grew up under the following conditions: The strain was incubated (5%, v/v) in 50 mL wort medium (pH 6.0) with 100 mg/L Fe ion and 120 mg/L Zn ion. The medium was loaded into a 250-mL Erlenmeyer flask and shaken in a rotary shaker (200 rpm) at 30°C for 60 h. Ferrous sulfate and zinc sulfate were chosen as the source of Fe and Zn. The Fe and Zn contents of the dry cells were determined by atomic absorption spectrum analysis. Under the optimized cultivation conditions, the Fe and Zn contents reached 7.854 mg/g dry cells and 4.976 mg/g dry cells.     

抗原抗体复合物型治疗性疫苗(乙克)临床研究中患者血清乙型肝炎病毒表面抗原下降的分析与启示

近年来全球慢性乙型肝炎(chronic hepatitis B,CHB)防治指南提出了“功能性治愈”(functional cure)的概念,即患者经过治疗达到血清乙型肝炎病毒表面抗原(hepatitis B virus surface antigen,HBsAg)消失,但现有抗病毒治疗很难实现这一目标。本研究对既往临床试验中经抗原抗体复合物型治疗性疫苗(乙克)治疗后的CHB患者HBsAg下降情况进行了归纳分析,结果显示,经乙克治疗随访后达到乙型肝炎e抗原(hepatitis B e antigen,HBeAg)血清学转换者的HBsAg下降高达0.95log₁₀IU/mL,显著高于未达到HBeAg血清学转换者的0.32log₁₀IU/mL(P<0.01),而经氢氧化铝佐剂治疗随访后发生HBeAg血清学转换(0.49log₁₀IU/mL)者与未发生HBeAg血清学转换者(0.36log₁₀IU/mL)之间HBsAg下降无统计学差异。乙克组治疗过程中,丙氨酸氨基转移酶(alanine aminotransferase,ALT)骤升(ALT flare)在HBsAg下降>1.0log₁₀IU/mL者中较多见,氢氧化铝组未观察到此现象。回归分析显示,乙克治疗后HBsAg下降的影响因素有患者出现HBeAg血清学转换、感染的HBV为B基因型、治疗过程中ALT出现10倍增高,以及基线血清HBsAg为高水平。结果提示,乙克诱导的特异性免疫对降低CHB患者血清HBsAg水平有一定效果,采用“抗病毒药物治疗＋针对HBsAg的中和性抗体被动免疫＋乙克主动免疫”的“三明治”治疗策略可能会提高“功能性治愈”率。