植物碱性亮氨酸拉链(bZIP)蛋白的研究进展(二)———DNA结合特性、基因表达、功能及应用

植物碱性亮氨酸拉链(bZIP)蛋白在高等植物基因表达与调控中起重要作用。本文介绍了植物bZIP蛋白与DNA结合特性,探讨了它们的基因表达和功能,综述了它们在分子生物学和基因工程研究中的应用。 Abstract:Plant basic leucine zipper (bZIP) proteins play an important role in the expression and regulation of higher plant genes.The DNA-binding properties of plant bZIP protein are introduced first in this article.Then their expression and function are discussed.Finally,their application to the studies of molecular biology and genetic engineering is reviewed.     

λ噬菌体穿孔素(holin) 蛋白触发裂菌的分子机制

穿孔素-裂解酶二元裂解系统是双链DNA噬菌体普遍采用的裂菌模式,以λ噬菌体为例,系统地揭示了噬菌体穿孔素的结构与功能。λ噬菌体的S基因的特征是呈双起始基序(dual-start motif),编码穿孔素(holin)S105和抗穿孔素(antiholin)S107,通过二者不同水平的表达及相互作用,触发裂菌过程。作者综述了λ噬菌体穿孔素的膜拓扑结构和成孔机制的最新研究进展,并展望了穿孔素的研究热点和应用前景。     

细菌核糖体的拯救机制

蛋白质翻译过程中,很多因素可能导致核糖体在mRNA上熄火,这对细胞的危害很大,因为这不仅占用了核糖体、氨基酸和tRNA,而且还可能产生有害的蛋白质.细菌进化出了多种核糖体拯救机制,释放熄火的核糖体,清除异常的mRNA,以规避毒害,如：① tmRNA·SmpB介导的拯救机制,又称为反式翻译介导的拯救机制|② ArfA(YhdL)介导的拯救机制|③ YaeJ(ArfB)介导的拯救机制.这些机制对于细菌的生理和繁殖都非常重要,但却在真核生物进化过程中消失了,使这些机制有可能成为抗菌药物靶点.本文主要就细菌核糖体的拯救机制做一概述,并对这些机制的应用前景进行了展望.     

噬菌体与细菌相互作用的分子机制研究进展

噬菌体与细菌是自然界中存在最广泛的两类微生物,两者在群体水平、个体水平以及分子水平上均存在复杂的相互作用关系.细菌能够影响溶原性噬菌体的溶原-裂解决策,而被噬菌体感染的细菌基因表达谱也会受到噬菌体影响,使宿主菌的代谢、应激、抵抗力、毒性等多种性状发生改变.现从细菌和噬菌体两者的角度,分别综述细菌抵抗噬菌体感染以及噬菌体...     

KLKs促进肿瘤增殖、迁移与侵袭的分子机制

组织激肽释放酶家族（kallikrein-related peptidases,KLKs）是一类具有胰蛋白酶或胰凝乳蛋白酶活性的分泌型丝氨酸蛋白酶,由15个成员（KLK1～15）组成。虽然发现较早,但关于其在肿瘤的发生与发展中所扮演的角色研究相对较少。KLKs作为肿瘤标志物在临床上的应用也有相应报道,如KLK3(PSA)作为前列腺特异性抗原用于前列腺癌的筛查。近年研究发现,KLKs家族在肿瘤进程中发挥重要作用,被认为是调节肿瘤细胞增殖、迁移和侵袭的关键因素。本文概述了在肿瘤进展过程中KLKs作用机制的最新进展,为其作为抗肿瘤治疗靶点的研究提供理论基础。     

光合细菌(Rhodopseudomonas capsulata)固氮活性与钼的吸收

利用紫色非硫细菌能在厌气光照下和好气黑暗下交替生长的特点和同位素~(99)Mo示踪,来探讨Rhodopseudomonas capsulata中Mo的积累与固氮酶合成的关系。 用硫酸铵和谷氨酸盐作为氮源,把Rps. capsulata置于厌气光照下生长。由于硫酸铵阻遏固氮酶,所以菌体内既无固氮酶活性也无~(99)Mo积累。而谷氨酸盐解遏固氮酶的合成,菌体则显示固氮活性并有~(99)Mo积累。 黑暗好气生长的Rps. capsulata菌体既无固氮活性,也没有~(99)Mo的积累。将这样的菌体转移到含~(99)Mo(无谷氨酸)的培养液进行光照,固氮酶活性迅速出现,同时有~(99)Mo的积累。在Rps. capsulata中钼的吸收与固氮酶的合成及活性是紧密偶联的。     

异化Fe(Ⅲ)还原酶促反应及调控机制的研究进展

异化Fe(Ⅲ)还原菌不是分类学上的概念,它具有系统发育及环境来源多样性的特点。与其他大多数的电子受体不同,在近中性pH值条件下,Fe(Ⅲ)的溶解度很低,通常以不溶性的Fe(Ⅲ)氧化物的形式存在。目前,对微生物如何获得和还原不溶性Fe(Ⅲ)的机理仍缺乏系统的了解。以希瓦氏菌和地杆菌为例,本文综述了3种异化Fe(Ⅲ)还原的酶促反应机制及其分子调控机理：异化Fe(Ⅲ)还原菌与Fe(Ⅲ)氧化物直接接触机制、电子穿梭体的作用机制、铁载体作用机制,多种膜蛋白特别是多血红素的细胞色素蛋白参与微生物的异化Fe(Ⅲ)还原过程,并形成复杂的调控网络。此外,本文也对异化Fe(Ⅲ)还原酶促反应及其分子调控机理将来的研究方向进行了展望,以期对这一重要的生化过程有更为全面的认识。     

AhR与Nrf2基因相互作用的分子机制研究进展

芳香烃受体(Aryl hydrocarbon receptor,AhR)作为一种配体依赖性激活的转录因子,调控肿瘤细胞的生长、增殖和凋亡,对环境中毒物和异物质代谢、机体免疫调节具有重要作用.核因子E2-相关因子2(Nuclear factor erythroid 2-related factor 2,Nrf2)是氧化还...     

海洋微生物裂解二甲基巯基丙酸内盐(DMSP)的酶促催化机制

二甲基巯基丙酸内盐(dimethylsulfoniopropionate,DMSP)是全球硫循环和碳循环的重要载体物质。海洋浮游植物、大型藻类和临海被子植物是DMSP的主要生产者。每年DMSP的产量可以达到1×10~9吨。在北大西洋表面的某些区域,DMSP的产量可以达到碳固定总量的10%。微生物介导的DMSP的裂解是全球硫循环和碳循环的重要步骤。目前,8种参与裂解DMSP的DMSP裂解酶已被报道。在已发现的8种DMSP裂解酶中,3种DMSP裂解酶的催化机制得到了研究和阐明。本文根据国内外研究成果,主要对DMSP裂解过程的酶促催化机制的研究进展进行综述,认为在今后工作中需要继续发现新的DMSP裂解酶,并进一步揭示海洋微生物裂解DMSP的分子机制。     

Molecular Mechanisms for Bacterial Potassium Homeostasis

Potassium ion homeostasis is essential for bacterial survival, playing roles in osmoregulation, pH homeostasis, regulation of protein synthesis, enzyme activation, membrane potential adjustment and electrical signaling. To accomplish such diverse physiological tasks, it is not surprising that a single bacterium typically encodes several potassium uptake and release systems. To understand the role each individual protein fulfills and how these proteins work in concert, it is important to identify the molecular details of their function. One needs to understand whether the systems transport ions actively or passively, and what mechanisms or ligands lead to the activation or inactivation of individual systems. Combining mechanistic information with knowledge about the physiology under different stress situations, such as osmostress, pH stress or nutrient limitation, one can identify the task of each system and deduce how they are coordinated with each other. By reviewing the general principles of bacterial membrane physiology and describing the molecular architecture and function of several bacterial K⁺-transporting systems, we aim to provide a framework for microbiologists studying bacterial potassium homeostasis and the many K⁺-translocating systems that are still poorly understood.     

Flexibility of the Bacterial Chaperone Trigger Factor in Microsecond-Timescale Molecular Dynamics Simulations

The bacterial chaperone trigger factor (TF) is the first chaperone to be encountered by a nascent protein chain as it emerges from the ribosome exit tunnel. Experimental results suggest that TF possesses considerable conformational flexibility, and in an attempt to provide an atomic-level view of this flexibility, we have performed independent 1.5-μs molecular dynamics simulations of TF in explicit solvent using two different simulation force fields (OPLS-AA/L and AMBER ff99SB-ILDN). Both simulations indicate that TF possesses tremendous flexibility, with huge excursions from the crystallographic conformation caused by reorientations of the protein’s constituent domains; both simulations also predict the formation of extensive contacts between TF’s PPIase domain and the Arm 1 domain that is involved in nascent-chain binding. In the OPLS simulation, however, TF rapidly settles into a very compact conformation that persists for at least 1 μs, whereas in the AMBER simulation, it remains highly dynamic; additional simulations in which the two force fields were swapped suggest that these differences are at least partly attributable to sampling issues. The simulation results provide potential rationalizations of a number of experimental observations regarding TF’s conformational behavior and have implications for using simulations to model TF’s function on translating ribosomes.     

Flexibility of the Bacterial Chaperone Trigger Factor in Microsecond-Timescale Molecular Dynamics Simulations

The bacterial chaperone trigger factor (TF) is the first chaperone to be encountered by a nascent protein chain as it emerges from the ribosome exit tunnel. Experimental results suggest that TF possesses considerable conformational flexibility, and in an attempt to provide an atomic-level view of this flexibility, we have performed independent 1.5-μs molecular dynamics simulations of TF in explicit solvent using two different simulation force fields (OPLS-AA/L and AMBER ff99SB-ILDN). Both simulations indicate that TF possesses tremendous flexibility, with huge excursions from the crystallographic conformation caused by reorientations of the protein’s constituent domains; both simulations also predict the formation of extensive contacts between TF’s PPIase domain and the Arm 1 domain that is involved in nascent-chain binding. In the OPLS simulation, however, TF rapidly settles into a very compact conformation that persists for at least 1 μs, whereas in the AMBER simulation, it remains highly dynamic; additional simulations in which the two force fields were swapped suggest that these differences are at least partly attributable to sampling issues. The simulation results provide potential rationalizations of a number of experimental observations regarding TF’s conformational behavior and have implications for using simulations to model TF’s function on translating ribosomes.     

A Three-protein Charge Zipper Stabilizes a Complex Modulating Bacterial Gene Silencing

The Hha/YmoA nucleoid-associated proteins help selectively silence horizontally acquired genetic material, including pathogenicity and antibiotic resistance genes and their maintenance in the absence of selective pressure. Members of the Hha family contribute to gene silencing by binding to the N-terminal dimerization domain of H-NS and modifying its selectivity. Hha-like proteins and the H-NS N-terminal domain are unusually rich in charged residues, and their interaction is mostly electrostatic-driven but, nonetheless, highly selective. The NMR-based structural model of the complex between Hha/YmoA and the H-NS N-terminal dimerization domain reveals that the origin of the selectivity is the formation of a three-protein charge zipper with interdigitated complementary charged residues from Hha and the two units of the H-NS dimer. The free form of YmoA shows collective microsecond-millisecond dynamics that can by measured by NMR relaxation dispersion experiments and shows a linear dependence with the salt concentration. The number of residues sensing the collective dynamics and the population of the minor form increased in the presence of H-NS. Additionally, a single residue mutation in YmoA (D43N) abolished H-NS binding and the dynamics of the apo-form, suggesting the dynamics and binding are functionally related.     

Bacterial Resistance to Nanosilver: Molecular Mechanisms and Possible Ways to Overcome them

This review contains the results of experimental studies of recent years dedicated to the resistance of bacteria to the action of nanosized silver and describes the putative molecular mechanisms of its development. Emphasis is placed on the study of works devoted to the investigation of the mechanisms of the resistance of bacteria to silver ions, which are the main factor of the bactericidal action of nanoparticles. The review also contains suggestions for further research aimed at developing of ways to overcome the problem of resistance of individual bacterial strains to the action of nanosilver and methods preventing its further spread.

     

InlA和InlB介导单核细胞增生李斯特菌入侵宿主细胞分子机制的研究进展

单核细胞增生李斯特菌(Listeria monocytogenes)是一种革兰氏阳性食源性致病菌。在造成宿主食源性感染的过程中, 单核细胞增生李斯特菌能凭借其独特的表面蛋白入侵宿主的非吞噬细胞。内化素蛋白家族(Internalins)是介导单核细胞增生李斯特菌入侵宿主非吞噬细胞的主要因子。本文根据国内外一些最新的研究成果, 结合作者近几年的工作, 综述了在侵染宿主的过程中, 单核细胞增生李斯特菌主要的内化素蛋白InlA和InlB介导细菌入侵宿主细胞的分子机制, 以期为阐明食源性致病菌致病机理、预防和治疗食源性疾病提供理论基础。