STE20 phosphorylation of AMPK-related kinases revealed by biochemical purifications combined with genetics

We have recently purified mammalian sterile 20 (STE20)–like kinase 3 (MST3) as a kinase for the multifunctional kinases, AMP-activated protein kinase–related kinases (ARKs). However, unresolved questions from this study, such as remaining phosphorylation activities following deletion of the Mst3 gene from human embryonic kidney cells and mice, led us to conclude that there were additional kinases for ARKs. Further purification recovered Ca²⁺/calmodulin-dependent protein kinase kinases 1 and 2 (CaMKK1 and 2), and a third round of purification revealed mitogen-activated protein kinase kinase kinase kinase 5 (MAP4K5) as potential kinases of ARKs. We then demonstrated that MST3 and MAP4K5, both belonging to the STE20-like kinase family, could phosphorylate all 14 ARKs both in vivo and in vitro. Further examination of all 28 STE20 kinases detected variable phosphorylation activity on AMP-activated protein kinase (AMPK) and the salt-inducible kinase 3 (SIK3). Taken together, our results have revealed novel relationships between STE20 kinases and ARKs, with potential physiological and pathological implications.     

SARS-CoV-2 envelope protein causes acute respiratory distress syndrome (ARDS)-like pathological damages and constitutes an antiviral target

Cytokine storm and multi-organ failure are the main causes of SARS-CoV-2-related death. However, the origin of excessive damages caused by SARS-CoV-2 remains largely unknown. Here we show that the SARS-CoV-2 envelope (2-E) protein alone is able to cause acute respiratory distress syndrome (ARDS)-like damages in vitro and in vivo. 2-E proteins were found to form a type of pH-sensitive cation channels in bilayer lipid membranes. As observed in SARS-CoV-2-infected cells, heterologous expression of 2-E channels induced rapid cell death in various susceptible cell types and robust secretion of cytokines and chemokines in macrophages. Intravenous administration of purified 2-E protein into mice caused ARDS-like pathological damages in lung and spleen. A dominant negative mutation lowering 2-E channel activity attenuated cell death and SARS-CoV-2 production. Newly identified channel inhibitors exhibited potent anti-SARS-CoV-2 activity and excellent cell protective activity in vitro and these activities were positively correlated with inhibition of 2-E channel. Importantly, prophylactic and therapeutic administration of the channel inhibitor effectively reduced both the viral load and secretion of inflammation cytokines in lungs of SARS-CoV-2-infected transgenic mice expressing human angiotensin-converting enzyme 2 (hACE-2). Our study supports that 2-E is a promising drug target against SARS-CoV-2.Subject terms: Cell death, Molecular biology     

合成己酸乙酯脂肪酶产生菌的筛选及产酶条件

从２７株脂肪酶产生菌中筛选到能由乙醇和己酸合成己酸乙酯的菌株８株。其中Ｒｈｉｚｏｐｕｓｓｐ．Ｈ－３菌株脂肪酶活力为５０－６０ｕ／ｍｌ,全细胞在有机溶剂中的酯化率可达己酸的９１％。Ｈ３产酶的最适碳源为淀粉或葡萄糖。６％黄豆饼粉加４％蛋白陈复合氮源有利于酶活力的增加。     

Selective pericentromeric heterochromatin dismantling caused by TP53 activation during senescence

Cellular senescence triggers various types of heterochromatin remodeling that contribute to aging. However, the age-related mechanisms that lead to these epigenetic alterations remain elusive. Here, we asked how two key aging hallmarks, telomere shortening and constitutive heterochromatin loss, are mechanistically connected during senescence. We show that, at the onset of senescence, pericentromeric heterochromatin is specifically dismantled consisting of chromatin decondensation, accumulation of DNA breakages, illegitimate recombination and loss of DNA. This process is caused by telomere shortening or genotoxic stress by a sequence of events starting from TP53-dependent downregulation of the telomere protective protein TRF2. The resulting loss of TRF2 at pericentromeres triggers DNA breaks activating ATM, which in turn leads to heterochromatin decondensation by releasing KAP1 and Lamin B1, recombination and satellite DNA excision found in the cytosol associated with cGAS. This TP53–TRF2 axis activates the interferon response and the formation of chromosome rearrangements when the cells escape the senescent growth arrest. Overall, these results reveal the role of TP53 as pericentromeric disassembler and define the basic principles of how a TP53-dependent senescence inducer hierarchically leads to selective pericentromeric dismantling through the downregulation of TRF2.     

发酵液内氨甲酰妥布拉霉素的定量测定

报道了应用一种以测量图象面积为指标的Ｚｙ－３００Ａ多功能抑菌圈测量仪,能够精确、稳定、快速地测定发酵液经薄层层析生物显迹后单组分氨甲酰妥布拉霉素的抑菌斑面积,然后从绘制的标准曲线中直接读取氨甲酸妥布拉霉素在发酵液中的含量,从而排除了其他组分对测定的干扰。试验证明,用Ｚｙ－３００Ａ多功能抑菌圈测量仪测定抗生素薄层层析生物显迹的抑菌斑,具有线性宽、精密度高、重复性强、操作简单方便等优点．本方法的建立为抗生素的纯品检定和多组分抗生素发酵、分离和纯化提供了准确的定量数据．     

烷化剂NTG诱变虾青素产生菌红法夫酵母的研究

虾青素是一种很有效的生物抗氧化剂和某些生物的天然着色剂,应用前景广阔。红法夫酵母是 生产虾青素的一个来源,优点颇多。天然菌株虾青素产量较少,缺少实用价值。实验采用烷化剂ＮＴＧ 诱变红法夫酵母,筛选出类胡萝卜素产量高的诱变株。用薄层层析对红法夫酵母产生的色素及其皂 化产物进行分析,并对各个成分的扫描光谱进行了比较。认为红法夫酵母产生的类胡萝卜素成分主 要是虾青素及虾青素二酯,还有一部分β－胡萝卜素。同时,还对虾青素产生的时相和ＢＨＴ对虾青素 光分解的保护作用进行了初步研究。     

松嫩平原羊草草地土壤-植物间铁的动态研究

羊草草地土壤-植物间营养元素Fe的动态研究结果表明,土壤中全铁和有效铁含量均较低,羊草中Fe含量比较丰富．在生长季各时期,全铁和有效铁含量在土壤剖面分布上从上向下呈下降趋势,有效铁与有机质含量呈显著正相关,而与pH值呈显著负相关．全铁各月平均含量变化在生长季呈“V”型曲线,7月含量最低;有效铁平均含量从5～8月与全铁相似,8月后则逐渐减少．羊草各器官及枯落物Fe含量有很大变化,总的趋势是根>根茎>叶>枯落物>茎;羊草叶、茎的Fe含量在生长季中为波动型下降曲线,根茎、根及枯落物的Fe含量在生长季中为“V”型曲线．A层土壤富集有效铁的能力稍强．土壤中Fe活性平均为0．640％。从5月至8月逐渐升高,8月后逐渐下降,10月份最低．地下部的Fe向地上部的转移强度为5～7月下降,8月升高,随后又下降．地上部Fe向枯落物的转移强度平均为105．0％,与地下部Fe向地上部转移强度呈显著负相关。     

江西武夷山中亚热带南方铁杉针阔混交林群落组成与结构

长期监测型固定样地是研究物种分布格局、群落动态和生物多样性维持机制等森林生态系统特征、过程与机理的重要平台。我国亚热带地区已建立的森林固定样地多为阔叶林类型,而针叶(阔叶混交)林类型十分有限。按照巴拿马Barro Colorado Island大型森林固定样地的建设规范,于2014年在江西武夷山国家级自然保护区内海拔1800 m左右的南方铁杉(Tsuga chinensis var. tchekiangensis)天然种群分布区域建立中亚热带针阔混交林6.4 hm~2固定样地。木本植物调查结果表明:(1)样地内胸径≥1 cm的木本植物共有29科53属89种,显著低于我国亚热带阔叶林样地平均水平,但显著高于温带针叶(阔叶混交)林样地平均水平;(2)在区系组成上,科以热带成分为主,而属以温带成分为主;(3)样地内独立个体数密度为2252株/hm~2,与温带针叶(阔叶混交)林平均水平相当,但显著低于亚热带阔叶林平均水平;(4)胸高断面积为37.89 m~2/hm~2,与亚热带阔叶林样地平均水平相当,但显著低于温带针叶(阔叶混交)林平均水平;(5)群落成层现象显著,优势种明显,多度排名前4位的物种的个体数占总个体数的55%,而排名后40的物种仅占1%;(6)群落总径级结构呈倒"J"型分布,其中胸径≤10 cm的小径木占总个体数的76.9%,而胸径30 cm的大径木仅占5.3%;(7)主要树种的径级结构有偏正态分布和"L"型分布等类型,但它们的种群在空间上均呈现显著的聚集分布特征;(8)南方铁杉虽然是群落现阶段最重要的优势种,但它的种群更新缓慢。研究结果充分显示该样地对中国森林多样性监测以及南方铁杉种群保育的重要意义。