hpc2研究进展

生物个体的胚胎发育以及细胞的增殖、分化,都同时受到多种基因的严格调控,PcG基因家族就是一类重要的发育相关基因.而hPc2基因是人PcG基因家族中的一个重要成员,其编码的HPC2蛋白,不仅可以和HPH、BMI-1以及RINGI等其他人类PcG蛋白结合形成HPC/HPH PcG复合体,以蛋白复合体的形式参与对homeotic基因的表达抑制,以维持机体的正常发育以及细胞的增殖和定向分化,还发现它能与其他多种蛋白质相结合,提示HPC2可能具有多种功能.因此,对hPc2的深入研究不仅有助于进一步阐明PcG基因家族的作用机理,扩展人们对基因表达调控的认识,还有助于发现PcG基因家族与其他信号转导通路的联系,更好地理解细胞信号网络系统.     

谷子PAL基因家族全基因组的鉴定和表达分析

苯丙氨酸解氢酶(phenylalanine ammonia-lyase,PAL)基因家族参与苯丙烷类代谢过程,通过调控植物抗病次生物质的合成在植物抗逆反应中发挥重要作用。为明确谷子PAL基因家族在逆境胁迫下的表达规律,该研究利用生物信息学方法对谷子PAL基因家族进行鉴定和表达分析。结果表明:谷子具有11个PAL基因,在进化树中可分为3个亚家族,SiPAL7独自进化为一支。通过构建蛋白结构域发现PAL基因家族成员均含有保守的PAL结构域。启动子分析显示,PAL基因含有应答激素、逆境胁迫等多种因子的顺式作用元件,说明PAL基因广泛参与不同生物学调控过程。RT-qPCR结果显示,谷子PAL基因家族多为诱导型表达,不同光照条件下PAL基因表达量变化明显,不同基因具有不同响应模式,说明谷子PAL基因家族在参与光调节反应中发挥重要作用。谷子PAL基因高度保守,广泛响应不同非生物胁迫,具有表达特异性。该研究结果为揭示PAL基因家族在调节谷子抗性及胁迫应答过程中的作用提供了参考。     

不同代次牛成肌细胞的诱导分化及相关基因的表达分析

利用初生荷斯坦牛的成肌细胞,在不同代次以含体积分数为2%马血清的DMEM进行诱导分化,在之后0、2、4、6、8、10 d观察细胞的形态变化并收集细胞提取总RNA,检测成肌相关基因的表达情况,为进一步研究牛肌肉发生过程及相关基因的表达调控提供依据。同时利用实时荧光定量PCR分别检测肌性相关基因MyoD(生肌决定因子)、MyoG(肌细胞生成素)以及非肌性相关基因A-FABP(脂肪细胞型脂肪酸结合蛋白)表达水平的变化。研究表明：①各代细胞在诱导培养4 d后开始有肌管形成,其后越来越多,8 d时达高峰。②成肌细胞向成熟肌细胞分化过程中,MyoD、MyoG、A-FABP基因都呈先上升然后下降的趋势。③高代次成肌细胞比低代次细胞增殖慢,而且在诱导分化后,肌管的数目明显变少; MyoD、MyoG、A-FABP随着代次的升高而下降。故推断MyoD、MyoG以及A-FABP基因会在成肌分化开始后的不同阶段被激活,从而发挥不同的调控作用,而且随着传代次数的增加成肌细胞的分化能力减弱。     

茶树TIFY基因家族鉴定及非生物胁迫下表达分析

TIFY基因家族在茶树激素信号转导及其逆境胁迫具有十分重要的作用。该文利用生物信息学方法对茶树基因组中的TIFY家族进行了鉴定,分析其理化性质、系统进化、基因结构、染色体定位、启动子区顺式作用元件、组织表达模式,并通过RT-qPCR对部分TIFY家族成员进行了非生物胁迫。结果表明:(1)茶树TIFY基因家族成员19个(CsTIFY1～CsTIFY19),分属于4个蛋白亚家族TIFY、JAZ、ZML 和 PPD,且不均匀地分布在8 条染色体上,按照进化关系及结构特点可分为 7 组,每组内具有相似的基因结构与保守基序组成。(2)CsTIFYs基因的启动子区具有多种包含激素和非生物胁迫响应的顺式作用元件,通过实时荧光定量(RT-qPCR)分析其家族成员对在茉莉酸甲酯、盐(20%氯化钠)、冷(4 ℃)以及干旱(20% PEG-6000)处理下反应强烈,部分基因在根与顶芽中有较高的表达量。由此推测,TIFY基因家族可能在茶树激素信号调控、胁迫响应、生长和发育等多方面发挥功能作用。     

同源盒基因(Hox)与哺乳动物生殖

哺乳动物的同源盒基因（Hox）与果蝇的同源异形基因是同源基因,该基因编码的DNA片段含183碱基对,转录由61个氨基酸残基组成的蛋白质保守结构域,称同源异型域.Hox基因碱基顺序及在染色体中的位置都是高度保守的.Hox基因在体节结构分化等空间信息调控中起着重要作用,按特异的空间模式赋予每一体节其自身的特点.近年来的研究表明,Hox基因不但影响胚胎发育,而且与成体生殖系统分化有关,在着床期子宫接受态的建立及子宫蜕膜反应的发生等生殖过程中起着重要的调节作用.     

肿瘤中neu基因的过量表达及其抑制

neu基因编码一种和表皮生长因子受体同源的磷酸蛋白,具有酪氨酸激酶的活性.近年来在多种人类肿瘤中发现neu基因的扩增和(或)过量表达.一些蛋白质因子或化学药物可以在转录水平阻遏neu基因的过量表达或者降低其产物p185^neu的酪氨酸激酶活性,抑制具有neu基因过量表达的癌细胞的转移和增殖.     

Neuronatin基因研究进展

Neuronatin(Nnat)是最近克隆的脑特异表达的新基因,在大脑发育过程中被选择性地剪接.人Nnat基因全长3 973 bp,含有3个外显子和2个内含子.Nnat mRNA有α和β两种剪接形式,分别编码81个和54个氨基酸,两者具有同一相位的开放阅读框架,差别在于β形式中间的外显子被剪切掉.人Nnat是单拷贝基因,定位于染色体20q11.2～12.小鼠Nnat基因定位于2号染色体末端.该基因是印记基因(imprinted gene),仅在父本来源的等位基因表达,而母本来源的等位基因则被甲基化不能表达.该基因在胚胎期及出生后早期大量表达,而在成年和衰老动物大脑中表达明显降低,因此推测该基因与大脑的发育和分化密切相关.成年动物中垂体前叶是Nnat mRNA唯一强表达的地方.推测其蛋白质产物具有疏水N端和亲水C端,是跨膜蛋白,其确切功能尚属未知.     

非编码的mRNA

非编码的mRNA是近年发现的一类不含典型ORF的mRNA.目前已发现或克隆的这类基因主要有：H19基因,XIST基因,XLSIRT基因,His-1基因,bic基因,rox1和rox2基因等.它们与胚胎发育,肿瘤发生及X染色体失活密切相关.     

甘蔗ScNRAMP基因家族的鉴定与生物信息学分析

NRAMP蛋白(natural resistance-associated macrophage proteins)家族在植物响应重金属胁迫时具有重要作用,能够转运Fe²⁺、Mn²⁺、Zn²⁺和Cd²⁺等重金属离子。为探究甘蔗ScNRAMP基因家族的特征,该文基于甘蔗割手密基因组鉴定了ScNRAMP基因家族,并进行了理化特性、基因结构、顺式作用元件、保守基序、结构域和进化关系等分析。结果表明:甘蔗ScNRAMP基因家族含有29个成员,不均匀分布在19条染色体上; 编码蛋白均为不稳定蛋白,无信号肽,亚细胞均定位在质膜上; 各成员保守基序有6～10个不等,跨膜数有6～12个不等,二级结构主要构成元件为α-螺旋和无规则卷曲; 顺式作用元件分析表明甘蔗ScNRAMP基因家族可能通过激素代谢参与逆境胁迫和生长发育等生物过程; 利用割手密的RNA-seq转录组表达数据进行的组织特异性分析发现,ScNRAMP在甘蔗不同发育阶段的叶和茎中具有时空表达特性; 进化树分析将甘蔗ScNRAMP家族成员分为3个亚家族(I、Ⅱ和Ⅲ)。该研究在全基因组水平上系统地鉴定了现代栽培甘蔗祖先种之一割手密NRAMP基因家族,既为进一步了解甘蔗NRAMP基因家族提供了基础,也为后续甘蔗重金属研究提供了重要候选基因。     

RNA干扰ABCE1基因后可增加肺癌95-D/NCI-H446细胞的E-钙黏附蛋白表达并减低细胞侵袭力

研究ABCE1对肺癌(95-D和 NCI-H446)细胞的作用．使用RNA干扰技术,抑制ABCE1基因的表达,通过Western blot 分析及FACS检测,观察ABCE1基因对E-钙黏附蛋白在95-D/NCI-H446细胞表达的影响;运用transwell 侵袭实验,观察M95-D/ NCI-H446细胞侵袭力的变化．RNA干扰ABCE1基因后,实验组与对照组相比,在48 h后可显著抑制肺癌(95-D和 NCI-H446)细胞ABCE1蛋白的表达,同时,伴随E-钙黏附蛋白的高表达,以及细胞侵袭力的降低． ABCE1基因与E-钙黏附蛋白相关,抑制ABCE1基因可增加肺癌95-D/NCI-H446细胞的E-钙黏附蛋白的表达,减低细胞的侵袭力．     

Selective interactions between vertebrate polycomb homologs and the SUV39H1 histone lysine methyltransferase suggest that histone H3-K9 methylation contributes to chromosomal targeting of Polycomb group proteins

Polycomb group (PcG) proteins form multimeric chromatin-associated protein complexes that are involved in heritable repression of gene activity. Two distinct human PcG complexes have been characterized. The EED/EZH2 PcG complex utilizes histone deacetylation to repress gene activity. The HPC/HPH PcG complex contains the HPH, RING1, BMI1, and HPC proteins. Here we show that vertebrate Polycomb homologs HPC2 and XPc2, but not M33/MPc1, interact with the histone lysine methyltransferase (HMTase) SUV39H1 both in vitro and in vivo. We further find that overexpression of SUV39H1 induces selective nuclear relocalization of HPC/HPH PcG proteins but not of the EED/EZH2 PcG proteins. This SUV39H1-dependent relocalization concentrates the HPC/HPH PcG proteins to the large pericentromeric heterochromatin domains (1q12) on human chromosome 1. Within these PcG domains we observe increased H3-K9 methylation. Finally, we show that H3-K9 HMTase activity is associated with endogenous HPC2. Our findings suggest a role for the SUV39H1 HMTase and histone H3-K9 methylation in the targeting of human HPC/HPH PcG proteins to modified chromatin structures.     

Characterization of Interactions between the Mammalian Polycomb-Group Proteins Enx1/EZH2 and EED Suggests the Existence of Different Mammalian Polycomb-Group Protein Complexes

In Drosophila melanogaster, the Polycomb-group (PcG) and trithorax-group (trxG) genes have been identified as repressors and activators, respectively, of gene expression. Both groups of genes are required for the stable transmission of gene expression patterns to progeny cells throughout development. Several lines of evidence suggest a functional interaction between the PcG and trxG proteins. For example, genetic evidence indicates that the enhancer of zeste [E(z)] gene can be considered both a PcG and a trxG gene. To better understand the molecular interactions in which the E(z) protein is involved, we performed a two-hybrid screen with Enx1/EZH2, a mammalian homolog of E(z), as the target. We report the identification of the human EED protein, which interacts with Enx1/EZH2. EED is the human homolog of eed, a murine PcG gene which has extensive homology with the Drosophila PcG gene extra sex combs (esc). Enx1/EZH2 and EED coimmunoprecipitate, indicating that they also interact in vivo. However, Enx1/EZH2 and EED do not coimmunoprecipitate with other human PcG proteins, such as HPC2 and BMI1. Furthermore, unlike HPC2 and BMI1, which colocalize in nuclear domains of U-2 OS osteosarcoma cells, Enx1/EZH2 and EED do not colocalize with HPC2 or BMI1. Our findings indicate that Enx1/EZH2 and EED are members of a class of PcG proteins that is distinct from previously described human PcG proteins.In Drosophila melanogaster, the genes of the Polycomb group (PcG) and trithorax group (trxG) are part of a cellular memory system, which is responsible for the stable inheritance of gene activity. The PcG and trxG genes have been identified in Drosophila as repressors (PcG) (18, 22, 27, 28, 38) and activators (trxG) (20, 21), respectively, of homeotic gene activity. PcG and trxG genes were originally found in Drosophila, but mammalian homologs have also been identified and appear to function like their Drosophila homologs (reviewed in reference 37). It has been proposed that PcG proteins repress gene expression through the formation of multimeric protein complexes. We have recently shown that the human PcG proteins HPH1 and HPH2 coimmunoprecipitate, cofractionate, and colocalize in nuclear domains with the human PcG proteins BMI1 (2, 12, 33) and HPC2, a recently identified, novel human Polycomb protein (33, 34). Furthermore, we have found that the human RING1 protein coimmunoprecipitates and colocalizes with HPC2 and other PcG proteins, indicating that RING1 is associated with, or is part of, the mammalian PcG complex (33, 35). These results indicate that mammalian PcG proteins form a multimeric protein complex. This observation is in agreement with observations that different PcG proteins, including Pc, bind in overlapping patterns on polytene chromosomes in Drosophila salivary gland cells (4, 10, 29).Interestingly, also the trithorax gene product trx colocalizes with Drosophila PcG proteins at many sites on polytene chromosomes (6, 24). Even more strikingly, binding of the trx protein has been mapped to small DNA fragments that also contain binding sites for PcG proteins, the Polycomb response elements (5, 6). This finding is further substantiated by the observation that GAGA factor, the gene product of the trxG gene trithorax-like (Trl) (13), colocalizes with Pc protein within the close vicinity of a Polycomb response element (41). Furthermore, the PcG gene Enhancer of zeste [E(z)] contains a domain with sequence homology with the activator protein trx (17). This observation is in agreement with genetic data which indicate that E(z) can be considered both a PcG gene and a trxG gene (26). Double mutations of E(z) and trxG genes result in homeotic phenotypes which are similar to the homeotic phenotypes which are also observed in double mutants of trxG genes (26). Finally, polytene chromosome binding of the trx protein is strongly reduced in homozygous E(z) mutants (4), and vice versa, polytene chromosome binding of the E(z) protein is reduced in trx mutants (24). These data suggest functional interactions between activators (trxG proteins) and repressors (PcG proteins) that are important for their mode of action.To start to investigate these puzzling features of the E(z) gene product, we used the two-hybrid system (8, 9) in order to identify proteins that interact with a mammalian homolog of E(z), the Enx1/EZH2 protein (15, 16). Here, we report the identification of the human EED protein, which interacts with Enx1/EZH2. EED is the human homolog of eed, a murine PcG gene (7, 36) which has extensive homology with the Drosophila PcG gene extra sex combs (esc) (14, 32, 39). Whereas Enx1/EZH2 and EED coimmunoprecipitate, they neither coimmunoprecipitate nor colocalize with other human PcG proteins, such as HPC2 and BMI1. Our findings indicate that both Enx1/EZH2 and EED form a class of mammalian PcG proteins that is distinct from previously described human PcG proteins.     

The pleiohomeotic functions as a negative regulator of <Emphasis Type="Italic">Drosophila even-skipped</Emphasis> gene during embryogenesis

Polycomb group (PcG) proteins maintain the spatial expression patterns of genes that are involved in cell-fate specification along the anterior-posterior (A/P) axis. This repression requires cis-acting silencers, which are called PcG response elements (PREs). One of the PcG proteins, Pleiohomeotic (Pho), which has a zinc finger DNA binding protein, plays a critical role in recruiting other PcG proteins to bind to PREs. In this study, we characterized the effects of a pho mutation on embryonic segmentation. pho maternal mutant embryos showed various segmental defects including pair-rule gene mutant patterns. Our results indicated that engrailed and even-skipped genes were misexpressed in pho mutant embryos, which caused embryonic segment defects.     

The dcr gene family of Desulfovibrio: Implications from the sequence of dcrH and phylogenetic comparison with other mcp genes

Desulfovibrio vulgaris Hildenborough contains a family of genes for methyl-accepting chemotaxis proteins (MCPs). Here we report the complete sequence of the gene for Desulfovibrio chemoreceptor H (dcrH). The deduced amino acid sequence of DcrH protein, which has an enlarged N-terminal, ligand binding domain, indicates a structure similar to that of other MCPs. Comparison of the sequences for DcrA, determined earlier, and DcrH indicated that similarity is essentially limited to the C-terminal excitation region. The dcr gene family differs, in this respect, from mcp gene families in other eubacteria (e.g. Escherichia coli and Bacillus subtilis), where MCPs share significant homology throughout their C-terminal signal transduction domains. This may point to an ancient evolutionary origin of the dcr gene family, which is widely distributed throughout the genus Desulfovibrio. The evolutionary origin of mcp genes was traced by comparing nucleotide sequences for the excitation region that is common to all MCPs. Phylogenetic analysis of sequences for thirty mcp genes from nine eubacterial and one archaebacterial species suggested that multiplication of mcp genes has occurred at least twice since the eubacteria diverged from the archaebacteria.Nucleotide accession number: The nucleotide sequence reported in this paper has been entered into GenBank under accession number U30319. Phone: 403-220-6388. Fax: 403-289-9311. Electronic mail address: voordouw@acs.ucalgary.ca.     

Effects of Polycomb Group Mutations on the Expression of Ultrabithorax in the Drosophila Visceral Mesoderm

The Polycomb group (PcG) genes encode repressors of many developmental regulatory genes including homeotic genes and are known to act by modifying chromatin structure through complex formation. We describe how Ultrabithorax (Ubx) expression is affected by the PcG mutants in the visceral mesoderm. Mutant embryos of the genes extra sex combs (esc), Polycomb (Pc), additional sex combs (Asx) and pleiohomeotic (pho) were examined. In each mutation, Ubx was ectopically expressed outside of their normal domains along the anterior-posterior axis in the visceral mesoderm, which is consistent with the effect of PcG proteins repressing the homeotic genes in other tissues. All of these four PcG mutations exhibit complete or partial lack of midgut constriction. However, two thirds of esc mutant embryos did not show Ubx expression in parasegment 7 (PS7). Even in the embryos showing ectopic Ubx expression, the level of Ubx expression in the PcG mutations was weaker than that in normal embryos. We suggest that in PcG mutations the ectopic Ubx expression is caused by lack of PcG repressor proteins, while the weaker or lack of Ubx expression is due to the repression of Ubx by Abd-B protein which is ectopically expressed in PcG mutations as well.     

毛竹APX基因系统进化与表达分析

抗坏血酸过氧化物酶(aseorbate peroxidase, APX)是植物活性氧代谢中重要的抗氧化酶之一,尤其是叶绿体中清除H₂O₂的关键酶,也是维生素C代谢的主要酶类。该文基于生物信息学方法,利用毛竹的基因组及转录组数据鉴定毛竹中的APX基因家族成员,并对其编码的蛋白基本理化性质、基因结构、启动子元件、系统进化及共线性关系、重复串联基因、GO注释及表达模式进行综合分析,共鉴定出21种编码APX的基因。结果表明:(1)PeAPX基因家族成员多为不稳定疏水蛋白,基因结构、基序及结构域相对较为保守,大多数APX基因具有高度保守的内含子模式。(2)系统进化关系显示毛竹APX基因与水稻APX基因有着较高的同源性关系,PeAPX具有较高的进化保守性。(3)Ka/Ks分析表明PeAPX基因都经历了纯化选择压力,此外在每个APX基因的启动子序列中发现有许多与应激反应和植物激素相关的顺式作用元件,结合表达量分析,表明毛竹APX基因在毛竹生长发育中起着正向促进作用。该研究为进一步了解毛竹APX基因家族基本功能及其抗氧化机制提供了一定的参考,为毛竹APX基因功能的深层次鉴定提供了重要依据。     

dBRWD3 Regulates Tissue Overgrowth and Ectopic Gene Expression Caused by Polycomb Group Mutations

To maintain a particular cell fate, a unique set of genes should be expressed while another set is repressed. One way to repress gene expression is through Polycomb group (PcG) proteins that compact chromatin into a silent configuration. In addition to cell fate maintenance, PcG proteins also maintain normal cell physiology, for example cell cycle. In the absence of PcG, ectopic activation of the PcG-repressed genes leads to developmental defects and malignant tumors. Little is known about the molecular nature of ectopic gene expression; especially what differentiates expression of a given gene in the orthotopic tissue (orthotopic expression) and the ectopic expression of the same gene due to PcG mutations. Here we present that ectopic gene expression in PcG mutant cells specifically requires dBRWD3, a negative regulator of HIRA/Yemanuclein (YEM)-mediated histone variant H3.3 deposition. dBRWD3 mutations suppress both the ectopic gene expression and aberrant tissue overgrowth in PcG mutants through a YEM-dependent mechanism. Our findings identified dBRWD3 as a critical regulator that is uniquely required for ectopic gene expression and aberrant tissue overgrowth caused by PcG mutations.     

Genetic and developmental analyses of chaetae pattern formation in Drosophila tergites

Summary The role of the achaete-scute complex and extramacrochaetae, Notch, Delta, Enhancer of split and Hairless genes in chaeta patterning in Drosophila tergites was studied in genetic mosaics and in mutant combinations. The mutant phenotypes of different alleles of each gene can be ordered in characteristic topographical seriations. These seriations are related to the pattern of proliferation of histoblasts and the time of singularization of sensory organ mother cells from surrounding epidermal cells. Genetic mosaics of lethal alleles show that these genes are fundamentally involved in this singularization and subsequent differentiation. The study of mutant combinations of alleles of these genes reveals specific relationships of epistasis and synergism between them. The results suggest that spatial and temporal variations in achaete-scute complex functional products in cells, modulated by the activity of other genes involved in signal transduction, define the patterned differentiation of sensory organs in tergites. Offprint requests to: A. García-Bellido     

CBL-CIPK信号系统参与小桐子抗冷性形成的生物信息学分析

类钙调磷酸酶B亚基蛋白(calcineurin B-like calcium sensor, CBL)属Ca²⁺结合蛋白,通过与类钙调磷酸酶B亚基互作蛋白激酶(calcineurin B-like calcium sensor interacting protein kinase, CIPK)互作介导Ca²⁺信号转导过程。CBL-CIPK信号系统参与了植物对多种逆境胁迫的响应过程。为深入探讨小桐子的抗冷性机制,该研究基于BLAST序列比对的方法,在全基因组水平对小桐子CBL与CIPK基因家族进行了鉴定,并对其系统进化、基因结构、表达特性及功能互作进行了解析。结果表明：(1)在小桐子基因组中共鉴定到8个CBL基因与18个CIPK基因,CBL与CIPK蛋白长度分别在211～257 aa与422～484 aa之间,等电点分别在4.65～5.08与6.20～9.26之间。(2)另外,CBL基因家族都包含8～10个外显子,而CIPK基因家族分为显著的1～2个外显子(11个基因)和12～15个外显子(7个基因)两类。(3)多序列比对显示,小桐子CBL蛋白...     

HDA19调控拟南芥对真菌Botrytis cineara的抗性反应

HDACs(Histone deacetylase)家族蛋白质负责组蛋白H3K4和H4K19脱乙酰化,并参与植物生长和应激反应的信号转导过程。茉莉酮酸酯Jasmonates(JA)是一种重要的天然植物激素,不仅调节植物生长和发育而且还参与植物对多种逆境胁迫响应的信号转导和调控过程。但是,HDACs在植物中参与JA信号转导的具体机制目前还不是很清楚。该研究以HDA19(Histone deacetylase 19)为对象,探讨了HDACs在植物JA信号转导中的功能和作用。结果表明:HDA19的T-DNA插入纯合突变体在JA处理条件下没有出现明显的JA根长反应。在相同处理条件下,hda19的不同突变体株系与相同生态型背景的野生型植株(WT)花色素苷含量无显著差异,但下游JAZ1、VSP1等JA信号通路的标记基因都显著上调表达。同时,hda19相比于WT对真菌Botrytis cineara的抗性显著增强,且hda19中下游基础防御标记基因PDF1.2、Thi2.1、ERF1等的表达水平显著高于WT。基于上述研究结果,该研究认为HAD19通过JA信号通路负调控拟南芥对真菌B.cineara的防御反应。