In Synechocystis sp. PCC 6803, gene sll1384 encodes a protein with a DnaJ domain at its N-terminal portion and a TPR domain at the C-terminal portion. An sll1384 mutant shows no difference from the wild type in adapta...In Synechocystis sp. PCC 6803, gene sll1384 encodes a protein with a DnaJ domain at its N-terminal portion and a TPR domain at the C-terminal portion. An sll1384 mutant shows no difference from the wild type in adaptation to different temperatures, but almost completely loses its capability of phototactic movement. After complementation with sll1384, the mutant regains the phototaxis. As shown with electron microscopy, on the cell surface, mutant cells have pili that appear to be the same as that of the wild type. Also, the transformation efficiency remains unchanged in the mutant. It is postulated that Sll1384 regulates phototaxis of Synechocystis through protein-protein interaction. It is the first DnaJ-like protein gene identified in a cyanobacterium for a role in phototaxis.展开更多
通过对大豆铝胁迫下的转录组测序分析,发现一个差异表达的基因,其编码一个具有101个氨基酸残基的Dna J-like分子伴侣蛋白-Gm Dna J1(Glycine max Dna J1),等电点为8.97;序列分析表明该蛋白具有典型的高度保守的J domain功能域,是一种类...通过对大豆铝胁迫下的转录组测序分析,发现一个差异表达的基因,其编码一个具有101个氨基酸残基的Dna J-like分子伴侣蛋白-Gm Dna J1(Glycine max Dna J1),等电点为8.97;序列分析表明该蛋白具有典型的高度保守的J domain功能域,是一种类型III的J蛋白;通过对其序列的同源性及进化关系分析,推测该蛋白可能响应重金属胁迫。为进一步探究Gm Dna J1是否能够对重金属胁迫产生应答反应,试验分别以0或100μmol·L-1Cu2+、Pb2+和Cd2+溶液胁迫处理的不同时间(0、12、24、48和72 h)大豆根尖RNA为材料,通过实时定量PCR研究了该蛋白基因的表达特征。结果表明:与对照相比,Gm Dna J1受Cu、Pb和Cd等重金属的诱导而强烈表达,呈现先升高后降低的趋势,其中Pb、Cd处理24 h后表达水平达到峰值,而Cu处理48 h后达到峰值;此外,Gm Dna J1对Cu、Pb和Cd胁迫的响应程度也不同,表明该基因对这三种重金属的响应模式存在差异。根据以上研究结果,推测大豆Gm Dna J1蛋白不仅响应铝毒胁迫,而且可能在响应重金属胁迫方面具有重要的作用,参与了大豆对重金属毒害的抵抗。该结果为深入研究Gm Dna J1在重金属胁迫响应中的功能及其分子机制提供了一定的依据。展开更多
Plants contain various factors that transiently interact with subunits or intermediates of the thylakoid multiprotein complexes, promoting their stable association and integration. Hence, assembly factors are essentia...Plants contain various factors that transiently interact with subunits or intermediates of the thylakoid multiprotein complexes, promoting their stable association and integration. Hence, assembly factors are essential for chloroplast development and the transition from heterotrophic to phototrophic growth. Snowy cotyledon 2 （SCO2） is a DNAJ-like protein involved in thylakoid membrane biogenesis and interacts with the light-harvesting chlorophyll-binding protein LHCBI. In Arabidopsis thaliana, SCO2 function was previ- ously reported to be restricted to cotyledons. Here we show that disruption of SC02 in Lotus japonicus results not only in paler cotyledons but also in variegated true leaves. Furthermore, smaller and pale- green true leaves can also be observed in A. thaliana sco2 （atsco2） mutants under short-day conditions. In both species, SCO2 is required for proper accumulation of PSlI-LHCll complexes. In contrast to other variegated mutants, inhibition of chloroplastic translation strongly affects L. japonicus sco2 mutant devel- opment and fails to suppress their variegated phenotype. Moreover, inactivation of the suppressor of variegation AtClpR1 in the atsco2 background results in an additive double-mutant phenotype with variegated true leaves. Taken together, our results indicate that SCO2 plays a distinct role in PSll assembly or repair and constitutes a novel factor involved in leaf variegation.展开更多
【目的】AsE246是我们首次报道的紫云英根瘤特异表达的非特异性转脂蛋白(nsLTP1:non specificlipid transfer protein 1)编码基因。本实验旨在筛选和鉴定与AsE246相互作用的宿主植物靶蛋白,并分析靶基因在共生和胁迫条件下的表达特征。...【目的】AsE246是我们首次报道的紫云英根瘤特异表达的非特异性转脂蛋白(nsLTP1:non specificlipid transfer protein 1)编码基因。本实验旨在筛选和鉴定与AsE246相互作用的宿主植物靶蛋白,并分析靶基因在共生和胁迫条件下的表达特征。【方法】利用酵母双杂交技术、小范围杂交技术及实时荧光定量PCR,筛选与AsE246的相互作用蛋白,并定量分析靶基因在结瘤与固氮过程中的时空表达特性。【结果】获取一个阳性克隆,其cDNA序列经Blast分析表明:候选靶蛋白是一个DnaJ-like蛋白,该蛋白相应基因命名为AsDJL1。AsE246与AsDJL1在酵母体内确实相互作用。AsDJL1在固氮根瘤中特异性增强表达,在NaCl胁迫下表达水平显著提高,在(NH4)2SO4胁迫下表达水平显著下降。【结论】本实验是筛选与LTP相互作用蛋白的首次报道。获得了直接的实验证据表明互作基因AsDJL1与AsE246具有高度相似的表达特征和功能,为深入研究二者的相互作用及其在共生固氮和应答环境胁迫中的调控机制,提供了一定的工作基础和理论依据。展开更多
基金Supported by the National Natural Science Fund for Distinguished Young Scientists (Grant No. 30825003)
文摘In Synechocystis sp. PCC 6803, gene sll1384 encodes a protein with a DnaJ domain at its N-terminal portion and a TPR domain at the C-terminal portion. An sll1384 mutant shows no difference from the wild type in adaptation to different temperatures, but almost completely loses its capability of phototactic movement. After complementation with sll1384, the mutant regains the phototaxis. As shown with electron microscopy, on the cell surface, mutant cells have pili that appear to be the same as that of the wild type. Also, the transformation efficiency remains unchanged in the mutant. It is postulated that Sll1384 regulates phototaxis of Synechocystis through protein-protein interaction. It is the first DnaJ-like protein gene identified in a cyanobacterium for a role in phototaxis.
文摘通过对大豆铝胁迫下的转录组测序分析,发现一个差异表达的基因,其编码一个具有101个氨基酸残基的Dna J-like分子伴侣蛋白-Gm Dna J1(Glycine max Dna J1),等电点为8.97;序列分析表明该蛋白具有典型的高度保守的J domain功能域,是一种类型III的J蛋白;通过对其序列的同源性及进化关系分析,推测该蛋白可能响应重金属胁迫。为进一步探究Gm Dna J1是否能够对重金属胁迫产生应答反应,试验分别以0或100μmol·L-1Cu2+、Pb2+和Cd2+溶液胁迫处理的不同时间(0、12、24、48和72 h)大豆根尖RNA为材料,通过实时定量PCR研究了该蛋白基因的表达特征。结果表明:与对照相比,Gm Dna J1受Cu、Pb和Cd等重金属的诱导而强烈表达,呈现先升高后降低的趋势,其中Pb、Cd处理24 h后表达水平达到峰值,而Cu处理48 h后达到峰值;此外,Gm Dna J1对Cu、Pb和Cd胁迫的响应程度也不同,表明该基因对这三种重金属的响应模式存在差异。根据以上研究结果,推测大豆Gm Dna J1蛋白不仅响应铝毒胁迫,而且可能在响应重金属胁迫方面具有重要的作用,参与了大豆对重金属毒害的抵抗。该结果为深入研究Gm Dna J1在重金属胁迫响应中的功能及其分子机制提供了一定的依据。
文摘Plants contain various factors that transiently interact with subunits or intermediates of the thylakoid multiprotein complexes, promoting their stable association and integration. Hence, assembly factors are essential for chloroplast development and the transition from heterotrophic to phototrophic growth. Snowy cotyledon 2 （SCO2） is a DNAJ-like protein involved in thylakoid membrane biogenesis and interacts with the light-harvesting chlorophyll-binding protein LHCBI. In Arabidopsis thaliana, SCO2 function was previ- ously reported to be restricted to cotyledons. Here we show that disruption of SC02 in Lotus japonicus results not only in paler cotyledons but also in variegated true leaves. Furthermore, smaller and pale- green true leaves can also be observed in A. thaliana sco2 （atsco2） mutants under short-day conditions. In both species, SCO2 is required for proper accumulation of PSlI-LHCll complexes. In contrast to other variegated mutants, inhibition of chloroplastic translation strongly affects L. japonicus sco2 mutant devel- opment and fails to suppress their variegated phenotype. Moreover, inactivation of the suppressor of variegation AtClpR1 in the atsco2 background results in an additive double-mutant phenotype with variegated true leaves. Taken together, our results indicate that SCO2 plays a distinct role in PSll assembly or repair and constitutes a novel factor involved in leaf variegation.
文摘【目的】AsE246是我们首次报道的紫云英根瘤特异表达的非特异性转脂蛋白(nsLTP1:non specificlipid transfer protein 1)编码基因。本实验旨在筛选和鉴定与AsE246相互作用的宿主植物靶蛋白,并分析靶基因在共生和胁迫条件下的表达特征。【方法】利用酵母双杂交技术、小范围杂交技术及实时荧光定量PCR,筛选与AsE246的相互作用蛋白,并定量分析靶基因在结瘤与固氮过程中的时空表达特性。【结果】获取一个阳性克隆,其cDNA序列经Blast分析表明:候选靶蛋白是一个DnaJ-like蛋白,该蛋白相应基因命名为AsDJL1。AsE246与AsDJL1在酵母体内确实相互作用。AsDJL1在固氮根瘤中特异性增强表达,在NaCl胁迫下表达水平显著提高,在(NH4)2SO4胁迫下表达水平显著下降。【结论】本实验是筛选与LTP相互作用蛋白的首次报道。获得了直接的实验证据表明互作基因AsDJL1与AsE246具有高度相似的表达特征和功能,为深入研究二者的相互作用及其在共生固氮和应答环境胁迫中的调控机制,提供了一定的工作基础和理论依据。
