TSA1 interacts with CSN1/CSN and may be functionally involved in Arabidopsis seedling development in darkness

The COP9 signalosome (CSN) is a multiprotein complex which participates in diverse cellular and developmental processes.CSN1,one of the subunits of CSN,is essential for assembly of the multiprotein complex via PCI (proteasome,COP9 signalosome and initiation factor 3) domain in the C-terminal half of CSN 1.However,the role of the N-terminal domain (NTD) of CSN 1,which is critical for the function of CSN,is not completely understood.Using a yeast two-hybrid (Y2H) screen,we found that the NTD of CSN1 interacts with TSK-associating protein 1 (TSA1),a reported Ca2+-binding protein.The interaction between CSN1 and TSA1 was confirmed by co-immunoprecipitation in Arabidopsis.tsal mutants exhibited a short hypocotyl phenotype in darkness but were similar to wild-type Arabidopsis under white light,which suggested that TSA1 might regulate Arabidopsis hypocotyl development in the dark.Furthermore,the expression of TSA1 was significantly lower in a csnl null mutant (fus6),while CSN1 expression did not change in a tsal mutant with weak TSA1 expression.Together,these findings suggest a functional relationship between TSA1 and CSN1 in seedling development.     

芥子气暴露的确证：家兔皮肤染毒后血红蛋白N端缬氨酸加合物的同位素稀释-NCI-GC/MS溯源性检测

素有＂毒剂之王＂之称的芥子气（HD）是目前危害最大的化学战剂之一,染毒后在体内可产生不同类型的特征性生物标志物,其对中毒诊断、溯源性分析以及毒理机制等研究有着重要的意义.本文首先采用同位素稀释-NCI-GC/MS分析方法监测和鉴定了HD体外全血染毒后产生的血红蛋白N端缬氨酸加合物（HETE-Val）;其次,研究了不同剂量HD（0.02～0.15LD50）经皮染毒家兔体内HETE-Val的时效、量效关系.结果表明,HETE-Val与家兔中毒剂量间有良好的量效关系,而时效关系表明染毒后15min内即可产生并检测到HETE-Val,一周内达到最大,随后逐渐下降,但至染毒后第103天仍可监测到该加合物.与血红蛋白N端缬氨酸加合反应的HD仅占HD染毒总量的～0.15‰,与家兔血液反应的HD约占HD染毒总量的～1%.表明HETE-Val是一种重要的生物标志物,并可应用于HD暴露的确证和化学武器核查的溯源性检测.     

肾功能与内环境恒定（二）

肾功能与内环境恒定（二）陈淑凤（北京师范大学生物学系１００８７５）（续１９９５年２９（１）：１８）３．２．１血液的缓冲系统血浆中的缓冲对为：碳酸（ＨＺＣＯ３）”蛋白质磷酸氢Ｍ钠（Ｎａ。ＨＰＯ。）磷酸Ｍ氢钠（ＮａＨｚＰＯ。）红细胞中的主要缓冲对为：一碳...     

WNK1： analysis of protein kinase structure, downstream targets, and potential roles in hypertension

The WNK kinases are a recently discovered family of serine-threonine kinases that have been shown to play an essential role in the regulation of electrolyte homeostasis, lntronic deletions in the WNK1 gene resuk in its overexpression and lead to pseudohypoaldosteronism type Ⅱ, a disease with salt-sensitive hypertension and hyperkalemia. This review focuses on the recent evidence elucidating the structure of the kinase domain of WNK1 and functions of these kinases in normal and disease physiology. Their functions have implications for understanding the biochemical mechanism that could lead to the retention or insertion of proteins in the plasma membrane. The WNK kinases may be able to influence ion homeostasis through its effects on synaptotagmin function.     

Prolonged Alzheimer-like tau hyperphosphorylation induced by simultaneous inhibition of phosphoinositol-3 kinase and protein kinase C in N2a cells

Co-injection of wortmannin (inhibitor of phosphatidylinositol-3 kinase, PI3K) and GF109203X(inhibitor of protein kinase C, PKC) into the rat brain was found to induce spatial memory deficiency and enhance tau hyperphosphorylation in the hippocampus of rat brain. To establish a cell model with durative Alzheimer-like tau hyperphosphorylation in this study, we treated N2a neuroblastoma cells with wortmannin and GF109203X separately and simultaneously, and measured the glycogen synthase kinase 3 (GSK-3)activity by y-32p-labeling and the level of tau phosphorylation by Western blotting. It was found that the application of wortmannin alone only transitorily increased the activity of GSK-3 (about 1 h) and the level of tau hyperphosphorylation at Ser^396/Ser^404 and Ser^199/Ser^202 sites (no longer than 3 h); however, a prolonged and intense activation of GSK-3 (over 12 h) and enhanced tau hyperphosphorylation (about 24 h) were observed when these two selective kinase inhibitors were applied together. We conclude that the simultaneous inhibition of PI3K and PKC can induce GSK-3 overactivation, and further strengthen and prolong the Alzheimerlike tau hyperphosphorylation in N2a cells, suggesting the establishment of a cell model with early pathological events of Alzheimer‘s disease.     

猪繁殖与呼吸综合征病毒N蛋白的原核表达、纯化与结构分析

将猪繁殖与呼吸综合征病毒 (PRRSV)BJ 4毒株N基因克隆至原核表达载体pET2 8a中 ,得到重组表达载体pET2 8 N ,转化EscherichiacoliBL2 1(DE3)细胞 ,获得可溶性表达 ,表达量占菌体蛋白的 2 8%。经ProbandNi2 亲和层析获得重组蛋白P2 8 N ,圆二色谱 (CD)测定结果表明 ,P2 8 N重组蛋白螺旋占 2 6 1% ,折叠占 2 3 7% ,转角 19 8% ,卷曲占 30 3%。并进一步绘制出PRRSVN蛋白的二级结构图     

防治禽流感特异性免疫球蛋白值得关注

中国工程院院士、军事医科院军事兽医研究所研究员夏咸柱近日表示,疫苗是禽流感最有效的预防方法,但由于病毒抗体漂移引起病毒的变异,生产应对当前流行株的疫苗至少需要4个月,这当然也包括H5N1抗原变异株。因此,在流感大流行的早期阶段应用抗病毒药物有助于控制疫情。WHO正准备利用现有的流感实验室网络,建立测定抗病毒药物敏感性检测机制,我国更有必要对国内的H5N1禽流感病毒株进行药物筛选与调查,为药物储备提供参考。     

中国学术期刊网络出版总库节点文献新疆驴脑组织尼帕病毒N基因片段的检测

目的检测新疆伊犁草原地区放养新疆驴的脑组织样本中尼帕病毒（Nipah Virus,NiV）核蛋白（N）基因片段,调查该地区放养新疆驴NiV感染流行状况。方法采用一步法实时荧光定量逆转录聚合酶链反应（one-step Real-Time FQ RT-PCR）对采自新疆伊犁地区草原放养且未接种NiV疫苗的65例新疆驴脑组织进行NiVN基因片段检测。结果新疆驴脑组织标本中未检出NiV N基因片段。结论目前尚无证据表明我国新疆伊犁地区新疆驴中存在NiV感染,提示该地区短时间内爆发该病毒感染可能性较小。     

利用靶定基质蛋白基因m的小干扰RNA抑制2009甲型H1N1流感病毒的复制

目的:设计、构建并筛选针对流感病毒基质蛋白基因m的小干扰RNA(siRNA),检测其对2009甲型H1N1流感病毒复制的抑制效果。方法:设计3条针对流感病毒m基因的siRNA,并克隆到短发夹型(shRNA)siRNA表达载体pSilencer2.1-U6-hygro上;经测序证明构建成功后,将表达3种siRNA的质粒和阴性对照质粒psiRNA-control分别转染MDCK细胞,用潮霉素筛选稳定表达细胞株,用H1N1流感病毒感染细胞,通过Real-time PCR和Western印迹检测干扰效果。结果:构建了3个针对流感病毒m基因的siRNA表达质粒,3种siRNA均使m基因的mRNA水平降低,其中M1-306的抑制效率达60%;3种siRNA均使流感病毒NA蛋白的表达量降低,M2-25、M1-105的抑制效率明显,M1-306略低。结论:针对m基因的siRNA可以有效抑制流感病毒在MDCK细胞中的复制。     

在无有机碳源和缺氧条件下自养菌和异养菌脱氨氮作用

在无有机碳源和缺氧条件下,应用^15N示踪考察4株具有氨氧化作用的菌株在膜反应器中的氨氧化产气情况。当溶氧浓度（DO）〈0．5mg／L的缺氧条件下,自养菌Nitrosomonas sp．单株培养,能将6．3％的氨氮转化为氮气,^15N2产生量占氨氮消耗量的21．86％。自养菌株与异养菌株混合培养,可将30．86％的氨氮转化为氮气,^15N2产生量占氨氮消耗茸的80．38％。企无有机碳的条件下,自养菌和异养菌的协同代时作用,可大大提高缺氧氩氧化产氮气的效率。