转录因子Atonal家族的研究进展

转录因子atonal因其具有一个碱性螺旋-环-螺旋（bHLH）结构而被归属于bHLH转录因子家族,在调节真核生物生长发育过程中起着重要作用.目前的实验研究已经证明它与视觉、听觉、嗅觉的形成有关,同时在不同时期还能够决定细胞的分化命运和抑制肿瘤的扩散.主要从模式生物果蝇和小鼠中的基因结构、组织分布、功能特征等方面研究概述,为进一步了解其他物种atonal转录因子的功能和作用机制奠定基础.     

MarR家族转录因子HpaR和XC0449协同调控野油菜黄单胞菌致病性

MarR家族转录因子广泛存在于细菌及古生菌中,并灵活、精细地调控多种毒力、抗胁迫及抗生素相关的生理生化途径。在野油菜黄单胞菌中,MarR家族转录因子HpaR (XC2827)的失活会显著降低细菌对于寄主甘蓝的致病力,同时会导致胞外蛋白酶的过量表达。本研究进一步发现,Xcc 8004基因组一共编码9个MarR家族转录因子。表达并纯化其中的HpaR (XC2827)和XC0449,体外微量热泳动(MST)实验及Pull-down实验证明二者可以在体外特异性结合。同时,表型检测发现XC0449突变会导致细菌致病力显著下降。通过体外凝胶迁移阻滞试验(EMSA)、体内qRT-PCR和GUS检测证明,XC0449和HpaR均作为转录激活子协同调控下游致病相关基因XC0705的表达,最终调控细菌毒力及胞外酶合成。     

Syntheses of some new methylcyclopentadienylmolybdenum complexes: Characterizations, crystal structures and comparisons with related complexes

As part of a long-term study of the substitution reactions of piano-stool type cyclopentadienylmetal carbonyl complexes, several new methylcyclopentadienylmolybdenum compounds have been prepared and characterized by methods including IR spectroscopy, electrospray ionization mass spectrometry and X-ray crystallography. The complexes reported here include [{Cp′Mo(CO)₃}₂I]BPh₄, cis-Cp′Mo(CO)₂(PPh₃)I and [Cp′Mo(CO)₃(CH₃CN)]BF₄ (Cp′ = η⁵-C₅H₄CH₃). In addition to their syntheses, comparisons are made between their IR spectroscopic and X-ray crystal structure data and those of similar complexes.     

水稻Trihelix转录因子家族全基因组分析及功能预测

Trihelix转录因子家族在植物生长发育以及响应逆境胁迫等方面发挥着重要作用,但目前基于水稻全基因组水平鉴定和分析该基因家族的研究尚未见相关报道。本文利用生物信息学方法在水稻基因组数据库中鉴定到Trihelix家族成员31个,序列聚类和功能结构域分析发现该家族均含有高度保守的、特征性的Trihelix结构域;根据亲缘关系远近和结构域特点,将其分为5个亚家族(Ⅰ~Ⅴ)。通过与拟南芥、二穗短炳草和高粱中Trihelix家族的聚类分析发现,这4个物种中Trihelix家族的分类相一致,但每个物种均含有不同亚家族的成员,表明该基因家族的分化早于物种的分化。基于MEME程序分析水稻Trihelix转录因子家族的保守基序与聚类分析结果具有较高的一致性。染色体区段复制分析表明,部分Trihelix家族成员在水稻以及水稻与其他物种之间存在种内和种间的染色体区段复制;生物芯片数据分析发现,Trihelix基因家族在水稻不同组织中、以及对6种不同植物激素的响应呈现多样化的表达谱。采用RiceFREND在线数据库分析发现,水稻Trihelix转录因子家族的20个成员与其他蛋白存在互作关系。本研究结果初步明确了水稻Trihelix转录因子家族的进化特点、染色体分布、染色体区段复制关系、组织表达、激素应答,以及该家族蛋白与其他蛋白质的互作情况,为进一步揭示Trihelix转录因子家族的分子进化规律和生物学功能奠定了基础。     

Synthesis and characterization of phosphorescent iridium complexes containing trifluoromethyl-substituted phenyl pyridine based ligands

Several iridium complexes containing trifluoromethyl-substituted phenyl pyridine based ligands have been synthesized and characterized to try to investigate the effect of trifluoromethyl group and its position on physical properties. The complexes have the general structure of (C-N)₂Ir(LX), where the C-N are 2-phenylpyridine (ppy), 2-(3,5-bis-trifluoromethylphenyl)pyridine (fmppy), 2-(3,5-bis-trifluoromethylphenyl)-4-methylpyridine (fmpmpy), 2-(3,5-bis-trifluoromethylphenyl)-5-trifluoromethylpyridine (tfmppy) and the LX are 2-picolinic acid (pic) and acetylacetonate (acac). The (tfmppy)₂Ir(pic) was characterized using X-ray crystallography. The absorption, emission, and thermostability of the complexes were systematically investigated. Introduction of CF₃ substituents into 2-phenylpyridine in (ppy)₂Ir(pic) lead to some decrease in the sublimation temperature, which is more suitable to devices fabrication. The experimental results revealed that the emissive colors of these complexes could be finely tuned by suitable incorporation of trifluoromethyl substituents on the 2-phenylpyridine ligand, obtaining bright green-blue emission λ_max values from 471 to 489 nm in CH₂Cl₂ solution at room temperature, with high solution quantum efficiencies ranging from 0.37 to 1.89 relative to Ir(ppy)₃.     

Alkali and alkaline-earth metal complexes of different nuclearities with azamacrocyclic pendant-arms ligands

A new series of alkali and alkaline-earth complexes of Na(I), Ca(II), Sr(II) and Ba(II) with two pyridyl pendant-arms azamacrocyclic ligands, L¹ and L², were synthesized by reaction of the ligand and the appropriate perchlorate salt in refluxing acetonitrile. The reactions gave analytically pure products that were characterized by elemental analysis, FAB mass spectrometry, IR, conductivity measurements and ¹H NMR spectroscopy. The crystal structure of [NaL¹](ClO₄), [BaL¹](ClO₄)₂ · 2CH₃CN and [Ca₂L²(μ-ClO₄)(H₂O)₂](ClO₄)₂ · Cl · (H₂O)_0.5 could be also determined. The crystal structure of the Na(I) and Ba(II) complexes with L¹ show endomacrocyclic mononuclear species, where the metal ions are in N8 and N10 coordination environment, respectively. The geometry around the metal ions can be described as cube and bicapped cube geometry for Na(I) and Ba(II), respectively. Instead, the crystal structure of the Ca(II) compound with L² shows an exomacrocyclic dinuclear complex where both metal ions are in a similar N5O2 environment joined through a bidentate perchlorate anion. The coordination geometry for both Ca(II) ions can be described as distorted pentagonal bipyramid.     

Sequence specific DNA binding of Ets-1 transcription factor: molecular dynamics study on the Ets domain--DNA complexes

Molecular dynamics (MD) simulations for Ets-1 ETS domain-DNA complexes were performed to investigate the mechanism of sequence-specific recognition of the GGAA DNA core by the ETS domain. Employing the crystal structure of the Ets-1 ETS domain-DNA complex as a starting structure we carried out MD simulations of: (i). the complex between Ets-1 ETS domain and a 14 base-pair DNA containing GGAA core sequence (ETS-GGAA); (ii). the complex between the ETS domain and a DNA having single base-pair mutation, GGAG sequence (ETS-GGAG); and (iii). the 14 base-pair DNA alone (GGAA). Comparative analyses of the MD structures of ETS-GGAA and ETS-GGAG reveal that the DNA bending angles and the ETS domain-DNA phosphate interactions are similar in these complexes. These results support that the GGAA core sequence is distinguished from the mutated GGAG sequence by a direct readout mechanism in the Ets-1 ETS domain-DNA complex. Further analyses of the direct contacts in the interface between the helix-3 region of Ets-1 and the major groove of the core DNA sequence clearly show that the highly conserved arginine residues, Arg391 and Arg394, play a critical role in binding to the GGAA core sequence. These arginine residues make bidentate contacts with the nucleobases of GG dinucleotides in GGAA core sequence. In ETS-GGAA, the hydroxyl group of Tyr395 is hydrogen bonded to N7 nitrogen of A(3) (the third adenosine in the GGAA core), while the hydroxyl group makes a contact with N4 nitrogen of C(4') (the complementary nucleotide of the fourth guanosine G(4) in the GGAG sequence) in the ETS-GGAG complex. We have found that this difference in behavior of Tyr395 results in the relatively large motion of helix-3 in the ETS-GGAG complex, causing the collapse of bidentate contacts between Arg391/Arg394 and the GG dinucleotides in the GGAG sequence.     

Synthesis and characterization of new lanthanide complexes with hexadentate hydrazonic ligands

In this paper, we report the synthesis and the characterization of a novel series of lanthanide (III) complexes with two potentially hexadentate ligands.The ligands contain a rigid phenanthroline moiety and two flexible hydrazonic arms with different donor atom sets (NNN′N′OO and NNN′N′N″N″, respectively for H₂L¹ (2,9-diformylphenanthroline)bis(benzoyl)hydrazone and H₂L² (2,9-diformylphenanthroline)bis(2-pyridyl)hydrazone).Both nitrate and acetate complexes of H₂L¹ with La, Eu, Gd, and Tb were prepared and fully characterized, and the X-ray crystal structure of the complex [Eu(HL¹)(CH₃ COO)₂] · 5H₂O is presented.The stability constants of the equilibria Ln³⁺ + H₂L¹ = [Ln(H₂L¹)]³⁺ and Ln³⁺ + (L¹)²⁻ = [Ln(L₁)]⁺ (Ln = La(III), Eu(III), Gd(III), and Tb(III)) are determined by UV spectrophotometric titrations in DMSO at t = 25 °C. The nitrate complexes of H₂L² with La, Eu, Gd and Tb were also synthesized, and the X-ray crystal structures of [La(H₂L²)(NO₃)₂(H₂O)](NO₃), [Eu(H₂L²)(NO₃)₂](NO₃) and [Tb(H₂ L²)(NO₃)₂](NO₃) are discussed.     

Construction and properties of pyruvate dehydrogenase complexes with up to nine lipoyl domains per lipoate acetyltransferase chain

The lipoate acetyltransferase (E2p) subunits of the pyruvate dehydrogenase (PDH) complex of Escherichia coli have three tandemly repeated lipoyl domains, although net deletions of one or two has no apparent effect on the activity of the purified complexes. Plasmids containing IPTG-inducible aceEF-lpd operons, which encode PDH complexes bearing from one to nine lipoyl domains per E2p chain (24-216 per complex), were constructed. They were all capable of restoring the nutritional lesion of a strain lacking PDH complex and they all expressed active sedimentable multienzyme complexes having a relatively normal range of subunit stoichiometries. The extra domains are presumed to protrude from the E2p core (24-mer) without significantly affecting the assembly of the E1p and E3 subunits on the respective edges and faces of the cubic core. However, the catalytic activities of the overproduced complexes containing four to nine lipoyl domains per E2p chain were lower than those with fewer lipoyl domains. This could be due to under-lipoylation of the domains participating in catalysis and interference from unlipoylated domains.     

互花米草NAC转录因子家族的鉴定与表达分析

互花米草(Spartina alterniflora)作为一种海岸带盐生植物,高度耐盐胁迫,但因为缺少参考基因组,其耐盐的分子机制却尚未见报道。NAC家族蛋白是植物特有的转录因子,调控植物的生长发育和胁迫应答。为了鉴定互花米草NAC蛋白(SaNAC)并探究它们与互花米草生长发育及胁迫响应之间的关系,本研究以互花米草三代全长转录组数据为参考,通过与水稻(Oryza sativa)、拟南芥(Arabidopsis thaliana)和玉米(Zea mays)NAC蛋白序列进行比对,并结合保守功能域进一步筛选,最终找到62个SaNAC蛋白。从蛋白序列比对、进化、motif预测、同源性比较、亚细胞定位、组织表达以及非生物胁迫下的基因差异表达等方面分别对互花米草NAC家族成员进行分析,结果发现SaNAC蛋白均含有保守的NAM结构域,且在进化上与水稻NAC家族具有一定的相似性;SaNAC家族中的两个蛋白SaNAC9和SaNAC49在细胞核表达;另外,本研究还发现互花米草SaNAC基因表达具有高度组织和胁迫应答差异性。这些结果表明互花米草NAC转录因子家族不仅具有保守的功能域,而且在调控互花米草的生长发育和非生物胁迫响应过程中具有重要的作用。     

Genome-wide identification,characterization and expression analysis of the LIM transcription factor family in quinoa

Lim family members play an important role in the regulation of plant cell development and stress response. However, there are few studies on LIM family in quinoa. In this study, we identified nine LIMS (named cqlim01-cqlim09) from quinoa, which were divided into three subfamilies (α Lim1, γ lim2 and δ lim2) according to phylogeny. The differences in gene structure and motif composition among different subfamilies have been observed. In addition, we studied the repetitive events of the members of the family. The Ka/Ks (non synchronous substitution rate / synchronous substitution rate) ratio analysis showed that the repetitive CqLIMs probably experienced purifying selection pressure. Promoter analysis showed that the family genes contained a variety of hormones, stress and tissue-specific expression elements, and protein interactions showed that these genes had actin stabilizing effect. In addition, QRT PCR results showed that all CqLIM genes were positively regulated under three stresses (low temperature, salt and ABA). These results provide a theoretical basis of further study of LIM gene in quinoa. Supplementary informationThe online version contains supplementary material available at 10.1007/s12298-021-00988-2.