Membrane Skeletal Proteins and Their Integral Membrane Protein Anchors are Targets for Tyrosine and Threonine Kinases in Euglena

ABSTRACT. Proteins of the membrane skeleton of Euglena gracilis were extensively phosphorylated in vivo and in vitro after incubation with [³²P]-orthophosphate or γ-[³²P] ATP. Endogenous protein threonine/serine activity phosphorylated the major membrane skeletal proteins (articulins) and the putative integral membrane protein (IP39) anchor for articulins. The latter was also the major target for endogenous protein tyrosine kinase activity. A cytoplasmic domain of IP39 was specifically phosphorylated, and removal of this domain with papain eliminated the radiolabeled phosphoamino acids and eliminated or radically shifted the PI of the multiple isoforms of IP39. In gel kinase assays IP39 autophosphorylated and a 25 kDa protein which does not autophosphorylate was identified as a threonine/serine (casein) kinase. Plasma membranes from the membrane skeletal protein complex contained threonine/serine (casein) kinase activity, and cross-linking experiments suggested that IP39 was the likely source for this membrane activity. pH optima, cation requirements and heparin sensitivity of the detergent solubilized membrane activity were determined. Together these results suggest that protein kinases may be important modulators of protein assembly and function of the membrane skeleton of these protistan cells.     

上海环城林带景观美学评价及优化策略

选取上海环城林带7种植物群落,采用美景度评判法,从林内景观和林外景观2个空间层次和春、夏、秋、冬4个季节,应用数量化理论Ⅰ建立了美景度和各景观因子类目之间的景观评价与预测的多元回归模型,分析了群落的结构特征和季相特征对林内景观以及外貌特征对林外景观的影响,并提出相应的优化对策。结果表明:(1)群落结构特征对林内景观的影响主要因子为胸径(平均胸径和胸径变异系数)、郁闭度和疏透度。在春季,林内美景度随着树木胸径增大而增加;在夏季,郁闭度增大会提升林内美景度;在秋季,胸径变异小的群落具有更高的林内观赏性;在冬季,疏透度对林内景观美景度影响最大。(2)群落季相特征对林内景观的影响,在各季节表现亦不同。在春季,黄色、紫色等明度较高的色相和开花量适中的群落美景度最佳;在夏季,生长势好、林冠层变化小以及树干清晰度高的群落具较高的美景度,且观花可显著提高夏季林内美景度;在秋季,色彩越纯美景度越高;而在冬季,树皮颜色深的群落美景度高。(3)群落外貌特征对林外景观有显著影响,其中林冠线对林外景观美景度影响最大,其次为林缘线。具有起伏不大林冠线和自然流畅林缘线的植物群落美景度高。旨在通过对典型植被群落不同季相的美景度评价,对上海环城林带的群落景观进行定量的评价,进而为不同情景下的群落结构优化提出相应的对策,为城市森林的群落建构与管理提供科学依据。     

河西走廊春季不同盐碱土壤中微生物数量、酶活性与理化因子的关系

【目的】揭示盐碱土壤微生物量与土壤因子间的关系。【方法】选择河西走廊不同盐碱程度的11个样点在春季进行采样,研究了土壤的微生物数量、酶活和理化性质,并对其进行方差分析、简单相关分析、逐步回归分析和主成分分析。【结果】河西地区原生盐碱地、次生盐碱地与农田土在土壤理化性质和土壤微生物数量等方面均有差异;河西地区土壤较贫瘠,土壤微生物数量较低,且分布有规律性,即原生盐碱土<次生盐碱土<农田土;放线菌、真菌、碱性磷酸酶、脲酶和有效磷5个因子是引起土壤微生物数量、酶活性与理化因子之间相关性的主要因素。【结论】结果证实河西地区盐碱土壤中磷的循环很大程度上影响着土壤微生物数量。     

小鼠淋巴细胞抗原CTLA-4的胞外肽段表达及纯化

目的: 真核细胞表达小鼠淋巴细胞抗原CTLA-4胞外段肽,研究表达肽段与抗原呈递细胞B7分子结合后减轻小鼠淋巴细胞刺激后的增殖抑制,从而启动T淋巴细胞进一步增殖。方法:从小鼠脾脏淋巴细胞获得总RNA,通过逆转录PCR扩增出CTLA-4全长基因,克隆并测序。依据胞外段序列和真核表达载体pcDNA3.1序列,合成引物扩增胞外片段,两者经内切核酸酶处理、连接构建重组表达pcDNA3.1载体,重组质粒经测序验证后,采用lipofectamine 2000转染入小鼠肝癌细胞Hepa1-6,经G418筛选获得稳定表达细胞株。结果:获得小鼠CTLA-4胞外段真核表达载体和小鼠肝癌细胞Hepa1-6稳定表达转染细胞株,制备了CTLA-4胞外肽段,经His标签抗体和小鼠CTLA-4抗体Western blot检测表达蛋白带均呈阳性。结论:获得CTLA-4胞外段肽,为进一步研究该肽的作用打下基础。     

Tumorigenic transformation of NIH 3T3 cells by the autocrine synthesis of transforming growth factor alpha

A variety of cancer cells overexpress transforming growth factor alpha (TGF alpha), a mitogenic peptide. A cDNA sequence coding for the full-length human TGF alpha precursor protein was subcloned into a retroviral expression vector and introduced into clone 7 NIH 3T3 cells, which have low numbers of endogenous epidermal growth factor receptors (EGFRs). The autocrine synthesis of TGF alpha by these cells resulted in their focal transformation. In contrast, control NIH 3T3 cells treated in a paracrine manner with exogenous, saturating concentrations of the mature form of TGF alpha, though stimulated to divide, remained morphologically untransformed. The addition of saturating quantities of soluble, mature TGF alpha to NIH 3T3 cells expressing the transferred TGF alpha gene actually suppressed their growth and focal transformation. The transformation induced by the TGF alpha gene remained an EGFR-dependent process, since the degree of transformation was correlated with EGFR expression in NIH 3T3 cells and since NR6 cells, which are Swiss 3T3 cells devoid of endogenous EGFRs, were transformed by the TGF alpha vector only when exogenous EGFR genes were also introduced. When inoculated into nude mice, the TGF alpha-expressing cells rapidly gave rise to tumors that grew progressively, whereas control cells did not form tumors. We conclude that in certain circumstances autocrine TGF alpha can be more oncogenic than paracrine and that paracrine TGF alpha can suppress this effect.     

中国植被分类系统修订方案

为了推动《中国植被志》研编工作, 该文回顾了中国植被分类系统的发展过程和主要阶段性成果, 提出了作为《中国植被志》研编技术框架组成部分的中国植被分类系统修订方案, 对各植被型组及各植被型进行了简单定义和描述, 并针对中国植被分类系统若干问题, 特别就中国植被分类系统总体框架、混交林的界定以及土壤在植被分类中的重要性等问题进行了讨论。1960年侯学煜在《中国的植被》中首次提出了中国植被分类的原则和系统, 1980年出版的《中国植被》制定了分类等级和划分依据等更加完善的系统, 之后《中国植被及其地理格局——中华人民共和国1:1 000 000植被图说明书》和《中国植物区系与植被地理》以及很多省区的植被专著对该系统进行过修订。2017年宋永昌在《植被生态学》中提出了一个分类等级单位调整的方案。本次提出的中国植被分类系统修订方案基本沿用《中国植被》的植被分类原则、分类单位及系统, 采用“植物群落学-生态学”分类原则, 主要以植物群落特征及其与环境的关系作为分类依据, 包含三级主要分类单位, 即植被型(高级单位)、群系(中级单位)和群丛(低级单位); 在三个主要分类单位之上分别增加辅助单位植被型组、群系组和群丛组, 在植被型和群系之下主要根据群落的生态差异和实际需要可再增加植被亚型或亚群系。修订方案包含了森林、灌丛、草本植被(草地)、荒漠、高山冻原与稀疏植被、沼泽与水生植被(湿地)、农业植被、城市植被和无植被地段9个植被型组, 划分为48个植被型(含30个自然植被型、12个农业植被型、5个城市植被型和无植被地段)。自然植被中有23个植被型进一步划分出了81个植被亚型。