钙调素通过HSP70调控热激转录因子的活性

凝胶阻滞分析结果显示,70kD的热激蛋白(HSP70)抑制热激转录因子(HSF)的DNA结合活性;利用免疫共沉淀法在细胞提取液中检测到了CaMHSP70复合物的存在,并且热激后CaMHSP70复合物含量增加.表明钙调素(CaM)调控HSF活性可能是通过与HSP70结合起作用.     

Cyclic AMP response element binding protein (CREB) participates in the heat-inducible expression of humanhsp90β gene

Human heat shock protein 90b gene ( hsp90b ) is a constitutively expressed heat shock gene existing in most of cell types tested that can be further induced by heat shock. Chloramphenical acetyl transferase (CAT) reporter plasmids driven by different regulatory fragments of hsp90b gene were constructed and transfected into Jurkat cells to explore the role of a cAMP response element (CRE) in the upstream of the gene. Results show that, in comparison with the wild type construct, a severe reduction (~2/3) in the increased folds of promoter activity induced by heat shock at 42℃ for 1 h was observed in a construct with CRE-containing fragment (-173/-91bp) deleted. Electrophoretic mobility shift assays (EMSA) showed that phosphorylated CRE-binding protein (CREB) in the nuclear extract of heat shocked Jurkat cells is specifically bound to the fragment. Additionally, both of the phosphorylation on CREB and the activity of protein kinase A (PKA) were found in Jurkat cells to be enhanced with extending time of heat shock treatment. Our results indicate that in addition to the intronic HSE/HSF pathway, phosphorylated CREB also participates in the heat shock induced expression of human hsp90b gene via its interaction with CRE which may be regulated by PKA-sig- naling pathway.     

真核生物的热击反应及转录调控的研究现状

真核生物受热击及其他应激作用后能诱导产生ＨＳＰ。ＨＳＰ在生物体内起着重要的生理功能。研究热击反应有助于搞清植物基因表达调控的机理，这也是科学家们近几年来研究的热点。随着植物基因工程技术的完善和发展，这一研究进展很快。ＨＳＥ、ＨＳＦ和ＨＳＰ在热击基因转录和调控过程中起着非常重要的作用。     

Activation in vitro of sequence-specific DNA binding by a human regulatory factor

The human heat-shock factor (HSF) regulates heat-shock genes in response to elevated temperature. When human cells are heated to 43 degrees C, HSF is modified post-translationally from a form that does not bind DNA to a form that binds to a specific sequence (the heat-shock element, HSE) found upstream of heat-shock genes. To investigate the transduction of the heat signal to HSF, and more generally, how mammalian cells respond at the molecular level to environmental stimuli, we have developed a cell-free system that exhibits heat-induced activation of human HSF in vitro. Comparison of HSF activation in vitro and in intact cells suggests that the response of human cells to heat shock involves at least two steps. First, an ATP-independent, heat-induced alteration of HSF allows it to bind the HSE; the temperature at which activation occurs in vitro implies that a human factor directly senses temperature. Second, HSF is phosphorylated. It is possible that similar multi-step activation mechanisms play a role in the response of eukaryotic cells to a variety of environmental stimuli, and that these mechanisms evolved to increase the range and flexibility of the response.     

水稻microRNA及其靶基因的系统鉴定

miRNA在植物生长发育和环境胁迫中发挥着重要的作用.本研究基于49个水稻小RNA 测序数据,系统分析了已知miRNA的表达,并鉴定到了新的水稻miRNA.我们发现,miRBase上注释的水稻miRNA中,只有39％具有可检测的表达. 此外,对水稻降解组数据的分析鉴定到一些新的miRNA靶基因,包括两个新的miR444靶基因,它们分别编码WD40蛋白和热休克因子型DNA结合蛋白.     

热激和热胁迫过程中玉米幼苗谷胱甘肽还原酶活性和同工酶的变化

 以玉米品种晴3和大黄为材料,研究结果表明热激幼苗的存活率比未热激幼苗高,这说明热激提高了玉米幼苗的抗热性.热激幼苗的谷胱甘肽还原酶活性明显高于未经热激的幼苗,同时热激处理后,GR的同工酶谱在整个热胁迫期间与对照相比也存在差异,说明热激过程中谷胱甘肽还原酶活性的升高和同工酶的变化与玉米幼苗的抗热性相关.     

黑腹果蝇热激蛋白基因家族的生物信息学分析

热激蛋白是细胞或生物体受到热激后新合成的一类遗传上高度保守的蛋白,在生物体中普遍存在,在细胞生长、发育、分化、基因转录等功能方面发挥重要的作用.本研究利用生物信息学方法首次对黑腹果蝇(Drosophila melanogaster)全部热激蛋白进行分析.结果表明,黑腹果蝇热激蛋白共有HSPC(HSP90)、HSPA(H...     

玉米幼苗热激诱导抗冷性过程中钙的效应

 玉米幼苗在冷胁迫前经过热激处理或CaCl₂浸种后再热激处理,其存活率,抗氧化酶GR的活性、可溶性蛋白质中热稳定蛋白质和游离脯氨酸的含量在的冷胁迫中均发生了变化,发现热激处理幼苗的这些参数高于对照,而最高的是CaCl₂浸种后再热激处理的,表明热激能提高玉米幼苗的抗冷性,而Ca²⁺对上述热激处理有加强作用.     

Activating protein factor binds in vitro to upstream control sequences in heat shock gene chromatin

DNA sequences, important for the control of Drosophila heat shock gene expression, are packaged in chromatin in a nuclease hypersensitive configuration. Recently, two protein-binding (exonuclease-resistant) sites which cover the TATA box sequence and an upstream control element were shown to occur in vivo amidst the 5' terminal hypersensitive regions of several heat shock genes. Protein-binding at the TATA box is independent of heat shock, but the binding at the upstream element is heat shock dependent, and it was proposed that a heat shock activator protein, HAP, positively regulates the genes. Here, I report the detection of HAP activity in heat shocked cell extracts by reconstituting specific binding to hsp82 gene chromatin in vitro. Inhibition of the binding by free DNA from the 5' region of heat shock genes implies a coordinate regulation of the gene family through HAP interaction with the upstream heat shock consensus sequence. Furthermore, the special ease of induction of the hsp82 gene over other heat shock genes can be explained in molecular terms by the higher affinity of HAP for the hsp82 binding site, which contains a 28 base sequence with almost perfect dyad symmetry, GAAGCCTCTAGAAG/TTTCTAGAGACTTC.     

Involvement of Ca<Superscript>2+</Superscript>/CaM in the signal transduction of acetylcholine regulating stomatal movement

It has been known that the neurotransmitter acetylcholine (ACh) also exists in plants and is able to regulate the movement of stomata.In another aspect,Ca^2 /CaM as the second messengers have a critical role of signal transduction in stomatal guard-cell,Here we showed that Ca^2 /CaM were also involved in the ACh regulated stomatal movement,In the medium containing Ca^2 ,the Ca^2 channel blockers (NIF and Ver)and CaM inhibitors (TFP and W7 ) could neutralize the ACh induced stomatal opening,however,they are ineffective in the medium containing K^ ,Those results indicated that Ca^2 /CaM were involved in the signal transduction pathway of ACh regulating stomatal movement.     

热诱导表达的水稻OsBBX30基因克隆和表达分析

生物信息学分析表明:OsBBX30基因启动子含有与逆境相关的作用元件HSE.为进一步了解OsBBX30基因在生物体内受热诱导,通过OsBBX30基因克隆构建原核表达和实时定量PCR分析,证实OsBBX30基因表达受热胁迫诱导,能增强大肠杆菌的耐热能力,为深入了解该家族基因和挖掘水稻耐热基因奠定基础.     

蚯蚓钙调素结合蛋白的研究

以赤子爱胜蚓(Eisenia Foetida)为材料,通过DEAE-Fast Flow离子交换层析、CaM-Sepharose亲和层析,分离纯化得到蚯蚓钙调素结合蛋白(CaMBPs)。纯化的CaMBPs对CaM激活的环核苷酸磷酸二酯酶活性有抑制作用,而且这种抑制作用可通过加入过量的CaM达到完全恢复。SDS-PAGE显示CaMBPs有3条明显主带,在EGTA存在时表现分子量分别为62, 49和30kD。紫外扫描测定含量分别为7.17%,7.31%和51.8%。用生物素-CaM覆盖法检测到3种CaM结合蛋白,与SDS-PAGE结果一致。酶活性测定实验表明在蚯蚓CaMBPs中有Ca²⁺-ATPase活性,但无NAD激酶活性。     

俄罗斯鲟雌核发育诱导及其后代的微卫星分析

通过冷休克和热休克法诱导2组俄罗斯鲟Acipenser gueldenstaedtii雌核发育,分别获得俄罗斯鲟的冷休克雌核发育组M1和热休克雌核发育组M2。利用6对具有高多态性的微卫星分子标记,分别对雌核发育系M1中的20尾鱼苗、M2中的40尾鱼苗、双亲及对照组20尾鱼苗基因组进行PCR扩增,并对结果进行基因型分析。分别得到2个雌核发育家系的期望杂合度(He)、等位基因数(A)和等位基因频率(P)。结果表明:冷休克雌核发育组的平均期望杂合度为0.591,平均等位基因数为6.0,等位基因频率为0.010～0.708;热休克雌核发育组的平均期望杂合度为0.687,平均等位基因数为5.5,等位基因频率为0.006～0.774。以父本特异微卫星条带作为诊断性标记,对雌核发育后代进行鉴定,结果发现在冷休克组和热休克组中分别存在40%和27.5%的杂交后代,此结果与表型鉴定结果相一致。另外,与母本基因型相比较,发现在冷休克组和热休克组中分别存在10%和15%的单倍体个体。只在热休克组中发现了完全的雌核发育后代,占总数的22.5%。冷休克和热休克雌核发育家系中除了单倍体后代,杂交后代和完全雌核发育后代外,还发现了不同程度的基因重组后代。