The N Termini of a-Subunit Isoforms Are Involved in Signaling between Vacuolar H+-ATPase (V-ATPase) and Cytohesin-2

Previously, we reported an acidification-dependent interaction of the endosomal vacuolar H⁺-ATPase (V-ATPase) with cytohesin-2, a GDP/GTP exchange factor (GEF), suggesting that it functions as a pH-sensing receptor. Here, we have studied the molecular mechanism of signaling between the V-ATPase, cytohesin-2, and Arf GTP-binding proteins. We found that part of the N-terminal cytosolic tail of the V-ATPase a2-subunit (a2N), corresponding to its first 17 amino acids (a2N(1–17)), potently modulates the enzymatic GDP/GTP exchange activity of cytohesin-2. Moreover, this peptide strongly inhibits GEF activity via direct interaction with the Sec7 domain of cytohesin-2. The structure of a2N(1–17) and its amino acids Phe⁵, Met¹⁰, and Gln¹⁴ involved in interaction with Sec7 domain were determined by NMR spectroscopy analysis. In silico docking experiments revealed that part of the V-ATPase formed by its a2N(1–17) epitope competes with the switch 2 region of Arf1 and Arf6 for binding to the Sec7 domain of cytohesin-2. The amino acid sequence alignment and GEF activity studies also uncovered the conserved character of signaling between all four (a1–a4) a-subunit isoforms of mammalian V-ATPase and cytohesin-2. Moreover, the conserved character of this phenomenon was also confirmed in experiments showing binding of mammalian cytohesin-2 to the intact yeast V-ATPase holo-complex. Thus, here we have uncovered an evolutionarily conserved function of the V-ATPase as a novel cytohesin-signaling receptor.     

Progress in the research of S-adenosyl-l-methionine production

This minireview mainly aims at the study of S-adenosyl-l-methionine (SAM) production by microbial fermentation. A brief introduction of the biological role and application of SAM was presented. In general, SAM production can be improved by breeding of the producing strain through the conventional mutation or genetic engineering approach in the molecular or cellular scale, by optimization of culture conditions in the cellular scale or bioreactor engineering scale, or by multiscale approach. The productivity of SAM fermentation has been improved greatly through the efforts of many researchers using the methods previously mentioned. The SAM-producing strains used extensively are Pichia pastoris and Saccharomyces cerevisiae. The effect of SAM on antibiotic production was also exemplified. The skill and scheme beneficial to the improvement of SAM production involves the enhancement of SAM synthetase (methionine adenosyltransferase) activity and selection of engineered constitutive promoters with appropriate strength; seeking for and eliminating the rate-limiting factors in SAM synthesis, namely, knocking off the genes that transform SAM and l-methionine (L-Met) to cysteine; release the feedback inhibition of SAM to methylenetetrahydrofolate reductase; blocking the transsulfuration pathway by interfering the responsible enzymes; enhancing ATP level through pulsed feeding of glycerol; and optimizing the L-Met feeding strategy. Precise control of gene expression and quantitative assessment of physiological parameters in engineered P. pastoris were highlighted. Finally, a discussion of the prospect of SAM production was presented.     

生长调节剂调控龙眼冲梢的研究

为高效、安全地调控龙眼冲梢,在龙眼花芽形态分化开始期(露红点期)和花穗主轴长6～9 cm的花穗展叶期,施用生长调节剂,比较了不同方法调控龙眼冲梢的效果。结果发现龙眼花芽形态分化开始期树冠喷施200 mg/kg乙烯利+150 mg/kg多效唑混合液和土施多效唑每株4 g处理控冲梢效果都显著高于对照,冲梢率分别是10.5%和7.6%,在花穗小叶处于展叶期虹吸输300 mg/kg乙烯利防控龙眼冲梢有效且安全。     

东海竹筴鱼的食性

以2008年5月、8月、11月和2009年2月东海灯光围网采集到的453条东海竹筴鱼为研究对象,对其胃含物进行分析,应用K-W非参数检验、卡方检验、聚类分析等方法,对不同季节和发育阶段条件下东海竹筴鱼的食性进行研究.结果表明: 东海竹筴鱼的饵料生物有124种(包括未鉴定种),浮游甲壳类和小型鱼类为其主要饵料类群.优势饵料生物依次是麦氏犀鳕(IRI%=39.2%)、长尾类糠虾幼体(IRI%=18.4%)、短尾类大眼幼体(IRI%=7.6%)和太平洋磷虾(IRI%=6.6%)等.季节和叉长对东海竹筴鱼的摄食强度均有显著影响(P<0.01),东海竹筴鱼春季摄食强度最高,而冬季最低;叉长140～159 mm的竹筴鱼摄食强度最高,叉长45~99 mm的幼鱼的摄食强度较高,其余叉长的鱼摄食强度相对较低.聚类分析结果表明,叉长100 mm是东海竹筴鱼摄食取向的拐点.东海竹筴鱼四季的平均营养级为3.51,属于低级肉食性鱼类.     

江西省弄蝶四新纪录(鳞翅目:弄蝶总科)(英文)

通过对外形特征和外生殖器特征的检视,发现江西省弄蝶四新纪录:无斑珂弄蝶指名亚种Caltoris bromus bromus(Leech,1894)、襟弄蝶中南亚种Pseudocoladenia dan fabia(Evans,1949)、灰陀弄蝶黎氏亚种Thoressa gupta leechii(Evans,1932)和南岭陀弄蝶Thoressa xiaoqingae Huang&Zhan,2004。标本均采自江西九连山国家级自然保护区。文中提供了成虫标本照片。     

CXCR4嗜性SHIVKU-1病毒静脉感染中国恒河猴初步研究

目的了解SHIVKU一1静脉途径感染中国恒河猴的感染特点及进展规律。方法两只健康中国恒河猴,静脉感染SHIVKU-1病毒,定期采样检测血浆病毒载量、CD4＋／CD8＋比值、CD4＋T细胞绝对数变化和血清中抗SHIVKU-1特异性IgG抗体水平。多色流式技术分析外周血、腹股沟淋巴结和十二指肠粘膜固有层CD4＋T淋巴细胞记忆细胞亚群变化。结果两只实验猴成功感染SHIVKU-1病毒,一直到感染后3个月均保持稳定水平的病毒载量。外周血CD4＋T淋巴细胞下降明显,CD4＋／CD8＋T细胞比值严重倒置。CD4＋Tcm细胞比例在经历了感染早期的下降后,大幅升高,尤其是外周血和淋巴结。CD4＋Tem则在粘膜固有层中增加明显。结论SHIVKU．1静脉途径成功感染了中国恒河猴,为SHIV／中国恒河猴疾病及评价模型的建立奠定了良好的基础,为今后使用此模型评价抗病毒药物或疫苗提供了条件。