Structural basis for the recognition of methylated histone H3 by the Arabidopsis LHP1 chromodomain

Arabidopsis LHP1 (LIKE HETEROCHROMATIN PROTEIN 1), a unique homolog of HP1 in Drosophila, plays important roles in plant development, growth, and architecture. In contrast to specific binding of the HP1 chromodomain to methylated H3K9 histone tails, the chromodomain of LHP1 has been shown to bind to both methylated H3K9 and H3K27 histone tails, and LHP1 carries out its function mainly via its interaction with these two epigenetic marks. However, the molecular mechanism for the recognition of methylated histone H3K9/27 by the LHP1 chromodomain is still unknown. In this study, we characterized the binding ability of LHP1 to histone H3K9 and H3K27 peptides and found that the chromodomain of LHP1 binds to histone H3K9me2/3 and H3K27me2/3 peptides with comparable affinities, although it exhibited no binding or weak binding to unmodified or monomethylated H3K9/K27 peptides. Our crystal structures of the LHP1 chromodomain in peptide-free and peptide-bound forms coupled with mutagenesis studies reveal that the chromodomain of LHP1 bears a slightly different chromodomain architecture and recognizes methylated H3K9 and H3K27 peptides via a hydrophobic clasp, similar to the chromodomains of human Polycomb proteins, which could not be explained only based on primary structure analysis. Our binding and structural studies of the LHP1 chromodomain illuminate a conserved ligand interaction mode between chromodomains of both animals and plants, and shed light on further functional study of the LHP1 protein.     

Pharmacological and genetic activation of cAMP synthesis disrupts cholesterol utilization in Mycobacterium tuberculosis

There is a growing appreciation for the idea that bacterial utilization of host-derived lipids, including cholesterol, supports Mycobacterium tuberculosis (Mtb) pathogenesis. This has generated interest in identifying novel antibiotics that can disrupt cholesterol utilization by Mtb in vivo. Here we identify a novel small molecule agonist (V-59) of the Mtb adenylyl cyclase Rv1625c, which stimulates 3’, 5’-cyclic adenosine monophosphate (cAMP) synthesis and inhibits cholesterol utilization by Mtb. Similarly, using a complementary genetic approach that induces bacterial cAMP synthesis independent of Rv1625c, we demonstrate that inducing cAMP synthesis is sufficient to inhibit cholesterol utilization in Mtb. Although the physiological roles of individual adenylyl cyclase enzymes in Mtb are largely unknown, here we demonstrate that the transmembrane region of Rv1625c is required during cholesterol metabolism. Finally, the pharmacokinetic properties of Rv1625c agonists have been optimized, producing an orally-available Rv1625c agonist that impairs Mtb pathogenesis in infected mice. Collectively, this work demonstrates a role for Rv1625c and cAMP signaling in controlling cholesterol metabolism in Mtb and establishes that cAMP signaling can be pharmacologically manipulated for the development of new antibiotic strategies.     

Lipid nanoparticle-encapsulated mRNA antibody provides long-term protection against SARS-CoV-2 in mice and hamsters

Monoclonal antibodies represent important weapons in our arsenal to against the COVID-19 pandemic. However, this potential is severely limited by the time-consuming process of developing effective antibodies and the relative high cost of manufacturing. Herein, we present a rapid and cost-effective lipid nanoparticle (LNP) encapsulated-mRNA platform for in vivo delivery of SARS-CoV-2 neutralization antibodies. Two mRNAs encoding the light and heavy chains of a potent SARS-CoV-2 neutralizing antibody HB27, which is currently being evaluated in clinical trials, were encapsulated into clinical grade LNP formulations (named as mRNA-HB27-LNP). In vivo characterization demonstrated that intravenous administration of mRNA-HB27-LNP in mice resulted in a longer circulating half-life compared with the original HB27 antibody in protein format. More importantly, a single prophylactic administration of mRNA-HB27-LNP provided protection against SARS-CoV-2 challenge in mice at 1, 7 and even 63 days post administration. In a close contact transmission model, prophylactic administration of mRNA-HB27-LNP prevented SARS-CoV-2 infection between hamsters in a dose-dependent manner. Overall, our results demonstrate a superior long-term protection against SARS-CoV-2 conferred by a single administration of this unique mRNA antibody, highlighting the potential of this universal platform for antibody-based disease prevention and therapy against COVID-19 as well as a variety of other infectious diseases.Subject terms: Biological techniques, Immunology     

ToxR负调控副溶血弧菌中c-di-GMP的合成

副溶血弧菌(Vibrio parahaemolyticus)是世界范围内引起海产品相关食物中毒的主要致病菌,具有很强的生物膜形成能力。ToxR是一种膜结合调控蛋白,对副溶血弧菌生物膜形成具有一定的调控作用,但具体机制尚未见报道。c-di-GMP是一种普遍存在于细菌中重要的第二信使,参与调控细菌的多种生物学行为包括生物膜的形成。本文探究ToxR对副溶血弧菌中c-di-GMP代谢的调控作用。利用酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)测定副溶血弧菌野生株(wild-type,WT)和toxR突变株(ΔtoxR)中c-di-GMP水平的差异。挑选c-di-GMP代谢相关基因scrA、scrG和vpa0198为进一步研究的靶标,采用实时定量qPCR实验检测靶基因在WT和ΔtoxR中的转录水平差异;将靶基因调控区DNA序列克隆入pHRP309质粒中无启动子的β半乳糖苷酶基因上游,采用lacZ报告基因融合实验进一步研究ToxR对靶基因的转录调控关系;将重组质粒分别导入含有pBAD33或pBAD33-toxR的EC100lpir中,采用lacZ报告基因融合实验研究ToxR是否能在异体宿主中调控靶基因的表达;PCR扩增靶基因上游调控区DNA序列,并纯化His-ToxR蛋白,用凝胶阻滞实验(electrophoresis mobility shift assay,EMSA)研究His-ToxR与靶基因启动子区DNA序列是否具有结合作用。ELISA结果显示ΔtoxR中c-di-GMP含量显著性高于WT中的,说明ToxR抑制c-di-GMP的产生;实时定量qPCR结果表明WT中scrA、scrG和vpa0198的转录水平显著性高于ΔtoxR中的,表明ToxR抑制它们的转录;lacZ报告基因融合实验结果表明ToxR可抑制副溶血弧菌和EC100lpir中scrA、scrG和vpa0198的启动子区活性;EMSA实验显示His-ToxR能特异性地结合到scrA和scrG的上游调控区DNA序列上,而对vpa0198的上游调控区DNA序列无结合作用。综上所述,ToxR通过直接调控相关酶蛋白基因的转录来抑制副溶血弧菌内c-di-GMP的合成,从而有助于精确调控生物膜形成等细菌行为。     

藏北申扎地区下石炭统永珠组下部孢子组合的特征及意义

报道藏北申扎永珠区德日昂玛—下拉剖面的永珠组孢子组合,发现孢子43属,70余种(包括未定种)。组合特征如下:1)缺乏欧美植物地理区早石炭世典型的石松类植物的小孢子Lycospora属及早石炭世晚期(维宪期)重要孢子属;2)出现较多早石炭世早期(杜内期),甚至泥盆纪晚期分子。呈现出新老化石的混合现象;3)北半球和南半球孢子的混合组合,同时也是冷、暖微古植物群混合,既有相当于中国早石炭世晚期大塘期以及西欧维宪期典型属种,又有南半球澳大利亚(包括巴西和非洲大陆)的维宪期的典型种。从孢子化石混合现象看,永珠组应为近岸浅水或滨海条件下的沉积,下伏地层的孢子化石再沉积现象说明当时有一次很大的海平面下降,在邻近地区出现了大面积的剥蚀区。说明杜内期末期—维宪期初期,申扎地区出现一次由冈瓦纳大陆冰川发育造成海平面相对下降。形成区域上不整合界面———洛工组或“巴日阿朗寨段”底部的不整合界面。混合孢子组合中的寒冷气候条件下的孢子化石的发现,印证了杨式溥、范影年和林宝玉推断的冈瓦纳冰川自早石炭世维宪期(大塘期)开始就影响到西藏申扎的推论。     

三种粘结剂用于纵折牙粘结的微渗漏研究

目的观察三种粘结材料的封闭性能,为折裂牙选择合适的粘结材料提供试验依据。方法选用健康前磨牙30颗,随机分为3组,制备成后牙纵折模型,分别用Clearfil SE Bond／AP—X（A组）、Super—Bond C＆B（B组）,Single Bond／F2000 Cmpomer（C组）进行粘结,在0．5％亚甲基蓝溶液中染色72h后,在电镜下观察染料渗透深度。结果扫描电镜下观察各组标本的粘结界面均有染料渗入,经单因素方差分析,A组与B组、C组有显著性差异,B组与C组之间无显著性差异。结论Clearfil SE Bond的封闭性能优于其它两组,但三种粘结剂均无法避免微渗漏的发生。     

中国海南内生拟盘多毛孢

2004 至 2005 年期间,从我国海南地区 95 种植物中分离鉴定了 43 种内生拟盘多毛孢,包括 5 个新组合,16 个内生拟盘多毛孢新记录种。5 个原归在盘多毛孢属 Pestalotia 的种重组到拟盘多毛孢属Pestalotiopsis 中,它们是二色拟盘多毛孢 P. bicolor,金鸡纳树拟盘多毛孢 P. cinchonae,密花拟盘多毛孢P. lambertiae,草原拟盘多毛孢 P. pampeana 和露兜树拟盘多毛孢 P. pandani。对 5 个新组合进行了重新描述和图解,其它 16 个内生拟盘多毛孢新记录种列出名录。