啮齿目核糖酸酶5基因(RNase5)的分子进化

RNase5是RNASE A基因超家族中的一个重要成员,是分子进化研究的理想模型之一。基于基因组水平,我们对啮齿目的3个进化枝10科17个物种开展RNase5的分子进化研究。利用TBlastN及BlastN方法鉴定每个基因组的RNase5基因,发现该基因在啮齿目的Ctenohystrica所有物种发生丢失,时间是在Ctenohystrica形成之后;邻接法和最大似然法构建的系统发育树均支持RNase5在“与小家鼠相关的进化枝”的小家鼠、褐家鼠和拉布拉多白足鼠发生三次独立基因复制事件;利用PAML软件的枝模型、位点模型及枝-位点模型计算选择压力,均检测到RNase5基因受到强烈的正选择作用。总之,我们的研究深入系统开展了RNase5在啮齿目中的分子进化,增加了该基因研究的多样性,为进一步系统认识该基因在动物的适应性进化遗传机制奠定了基础。     

植物查尔酮合成酶超基因家族的分子进化

查尔酮合成酶(CHS)超基因家族又称为植物类型III聚酮合酶超基因家族, 其编码酶通过催化和合成一系列结构多样及生理活性各异的次生代谢物, 在植物生长发育和适应环境的过程中扮演着重要角色。为全面了解CHS超基因家族在植物中的进化规律, 重建其进化历史, 该研究利用14种具有全基因组数据的代表植物, 通过生物信息学手段, 深入挖掘和分析了不同植物类群基因组中查尔酮合成酶超基因家族的成员构成, 推测了其可能的扩增机制和功能分歧, 并探讨了该超基因家族在植物中的总体进化趋势。结果共识别144条具有表达信息的同源序列, 它们全部来自9种陆生植物的基因组, 藻类植物基因组中没有发现相关序列。系统发育和进化分析表明, CHS超基因家族的起源古老, 它们可能为适应复杂的生态环境而出现在早期的陆生植物中, 之后在长期的进化过程中不断发生谱系的特异扩张和拷贝丢失, 最后通过功能分歧的形式在不同植物类群中被分别固定。此外, 进化检验也显示, 尽管CHS超基因家族内部发生了多样的遗传改变, 但整个超基因家族仍处于强烈的纯化选择之下, 并且个体基因中也无任何单氨基酸位点受到正向选择的影响。     

脊椎动物ABCA基因亚家族研究进展

ABC（ATP-binding cassette）基因家族编码膜蛋白,其成员负责多种物质的跨膜运输。基于氨基酸序列的同源性,人的48个ABC成员被分为7个亚家族：ABCA~ABCG。与其他亚家族相比,ABCA基因编码的蛋白具有独特的拓扑结构,并且其家族成员在两栖动物和哺乳动物分化之后各发生过一次大的扩展（expanding）。基因结构分析发现这两次扩展均是通过基因倍增实现的,这些倍增的产物在啮齿目和食肉目中得到保留,而在灵长目中却有一半变成假基因或被删除。ABCA成员主要负责不同组织器官脂类和胆固醇的跨膜运输,部分成员的突变与疾病相关。     

基于基因家族大小的比较研究脊椎动物的适应性进化

同源基因家族的拷贝数在不同物种间普遍存在差异,这种差异是由不同的基因得失速率引起。众所周知,基因拷贝数变异是特定物种表型创新的可能原因。本研究选取具有代表性的脊椎动物主要类群并跨约6亿年进化时间的64个物种,鉴定了它们的同源基因家族,揭示了脊椎动物基因家族大小的进化模式。结果表明:在推断的存在于脊椎动物最近共同祖先的6857个基因家族中,有6712个都在至少一个种系中发生了大小的变化,而且基因家族在大多数种系中都是收缩的;其中,霍氏树懒(Choloepus hoffmanni)中有最高的基因家族收缩水平,而在斑马鱼(Danio rerio)中则相反。基于脊椎动物基因家族大小进化的高度动态性,本研究从基因家族大小变化的角度鉴定了一些可能与特定脊椎动物类群进化有关的基因组信号。结果观察到在现存真骨鱼类最近共同祖先基因组中出现了可能因全基因组复制所导致的高比例的基因家族扩增现象,随后在后裔物种中发生基因收缩事件。此外,本研究还发现了硬骨鱼特异性的orphan基因可能对这些鱼类在水生环境中的适应性进化有所贡献的证据,如在有些硬骨鱼中orphan基因与鳍、尾巴、肾脏等发育有关。本研究结果有助于深入了解脊椎动物基因家族大小的进化,同时为理解脊椎动物基因组进化与表型多样性的联系提供了理论证据。     

基于全基因组的龟鳖目Hox基因序列特征及进化分析

为对Hox基因在龟鳖目物种中进行系统地序列比较分析和进化研究，文章对目前具有染色体水平的龟鳖目基因组进行了Hox基因的鉴定,序列特征、进化和转录组分析。研究结果表明龟鳖物种的Hox基因簇是高度保守的。非重复序列的缺失导致鳖科HoxB9—HoxB13基因间区相对龟科短了10 kb。大量Hox基因编码区发生了鳖科或龟科特异的序列替换、插入和缺失。胸部骨骼发育相关的Hox基因在鳖科祖先发生了快速进化和受到正选择。Hox基因的表达具有组织、时期特异性,主要在胚胎时期的顶端外胚层嵴、背甲嵴和性腺表达。研究为龟鳖目Hox基因不同胚胎时期的多组学及表达调控分析提供了靶标,也为进一步厘清龟鳖物种演化创新提供了参考。     

蒺藜苜蓿全基因组中WRKY转录因子的鉴定与分析

WRKY基因家族是植物基因组内一类重要的转录因子, 参与植物许多生理生化过程, 如植物发育、代谢以及生物和非生物胁迫。目前, 已在多种植物中鉴定出WRKY基因家族, 但是关于蒺藜苜蓿(Medicago truncatula L.) WRKY基因家族的系统分析鲜有报道。文章利用生物信息学方法, 从蒺藜苜蓿全基因组中共鉴定出93个WRKY基因, 包括81个标准的WRKY基因(19个Ⅰ型基因, 49个Ⅱ型基因以及13个Ⅲ型基因)和12个非标准类型的WRKY基因。对这些WRKY基因进行了基因重复、染色体定位、基因结构、保守基序和系统进化等方面的分析。蒺藜苜蓿WRKY基因家族中最近共发生了11次基因重复事件, 共涉及24个基因, 占全部WRKY基因的26%。染色体物理定位分析表明, 蒺藜苜蓿WRKY基因在染色体上呈不均匀分布, 存在6个基因簇。对WRKY Ⅲ型基因的进化分析表明, 它们在长期的进化过程中受纯化选择压力。     

真骨鱼类DMRT基因家族的连锁结构及其系统发生

利用已经报道的多个物种基因组序列资源,我们通过比较真骨鱼类（Teleost fish）共有的DMRT1-5基因所在染色体位置的基因组结构特征,从基因组水平证明了真骨鱼类和四足动物的相应DMRT基因的直系同源关系,揭示了DMRT4和DMRT5是因为真骨鱼类和四足动物的共同祖先发生了染色体重复事件而由同一基因分歧演变形成的。同时通过基因连锁分析,探测到包括SNF2和elavL家族基因在内的多个可能与DMRT基因功能密切相关的基因,为进一步研究DMRT基因和它们之间可能存在的调控机制提供了新的线索。     

牛科动物HSL基因序列分析及其分子进化研究

在对牛科中4种动物即牦牛、瘤牛、普通牛和水牛HSL基因外显子Ⅰ部分核苷酸序列进行测定的基础上,与Gen-Bank中其他物种相应基因核苷酸序列、氨基酸序列进行了比对分析,并构建了牦牛与其他物种间分子系统进化树。结果表明:牦牛与普通牛、瘤牛、水牛、猪、人、小鼠、大鼠7个物种HSL基因外显子Ⅰ部分核苷酸序列间保守性较高,同源性大小依次为99.8%、99.6%、97.4%、90.6%、88.4%、83.5%、82.3%。相应氨基酸序列间保守性更高,同源性分别为100%、100%、98.2%、94.0%、92.2%、89.8%、89.8%。牦牛与各物种该基因部分核苷酸序列间碱基变异类型主要表现为碱基转换和颠换,无碱基插入和缺失发生,碱基转换的频率高于颠换的频率;在核苷酸水平上的多数碱基替换都是同义替换;序列间单碱基变异位点大多出现在同一位点,多发生在密码子第3位,其次是第1位,最少发生在第2位,符合分子进化的中性学说。HSL基因外显子Ⅰ部分核苷酸序列进行多序列对位排列构建的各物种间分子系统进化树结果表明,普通牛和瘤牛首先聚为一类,再分别与牦牛、水牛、猪、人聚类,最后与大鼠、小鼠聚为一类。该聚类结果与动物学上的分类结果一致,表明HSL基因外显子Ⅰ部分核苷酸序列适合于构建物种间分子系统进化树。研究表明,牦牛、普通牛和瘤牛3个物种间的遗传距离大小相近,牦牛和水牛间的遗传距离与普通牛、瘤牛和水牛间的遗传距离大小相当。牦牛、普通牛和瘤牛3个物种间的遗传距离远小于它们各自与水牛这一物种的遗传距离,它们三者之间的亲缘关系也相对于它们各自与水牛间的亲缘关系都较近,故将牦牛、普通牛和瘤牛划分在同一个属——牛属(Bos)更为合理。     

miRNA-9基因家族进化分析及其靶基因预测

microRNAs(miRNA)是真核生物中一类长度约为21～25个核苷酸的非编码小分子RNA,在转录后水平调控基因的表达。该文在miRBase中搜索后生动物的mir-9基因序列。47个物种中共搜索到120条mir-9基因序列,说明mir-9基因家族广泛存在于不同物种中。基因定位显示86%的mir-9基因存在于基因间隔区(IGR),多序列比对发现miR-9基因家族成熟序列的第2位到第8位碱基以及第14位到第18位碱基为保守碱基。进化分析表明mir-9b和mir-9c可能是此基因家族最早出现的基因形式,即祖先基因。这些祖先基因经过串联重复、大片段重复、个别碱基的缺失及突变等方式形成了脊椎动物中miR-9-1至miR-9-7数个基因。分别采用四个miRNA靶基因预测软件对mmu-miR-9的靶基因进行预测,发现miR-9与神经系统发育、心肌系统疾病和跨膜运输系统等密切相关。该研究为今后进一步研究miRNA调控的神经系统发生和神经细胞生长与分化的机制奠定了基础。     

松鸡科13种鸟类COⅠ序列变异及系统发育分析

线粒体DNA中的细胞色素C氧化酶亚基Ⅰ基因,即COⅠ是DNA条形编码的主要基因,是一个很好的物种鉴定工具,目前广泛应用于鸟类系统发育研究。通过测定花尾榛鸡和黑琴鸡COⅠ基因的序列,并结合GenBank中松鸡科13种鸟类的同源序列,对松鸡科鸟类进行了序列变异和系统发育分析。结果显示,松鸡科物种的种间变异大于种内变异。序列分歧和系统分析结果支持花尾榛鸡Tetrastes bonasia归于松鸡科Tetraonidae榛鸡属Tetrastes。黑琴鸡Lyrurus tetrix与Tetrao属中其它物种的分歧小于松鸡科其它属间分歧,且黑琴鸡聚在Tetrao内,研究结果倾向支持黑琴鸡归于松鸡属Tetrao。     

Molecular Evolution of Prolactin Gene Family in Rodents

In this study, we identified two novel members of prolactin gene family in rat by blast searches against the published genomic database. A further analysis showed that gene duplications leading to PRL gene family in rodents occurred after rodents diverged from other mammals. Major reorganization of the gene loci in rodents was largely completed before the split of rat and mouse. But PL-I and PL-II genes are the exceptions, which have clustered in a species-specific manner in the phylogenetic tree. By combining results from gene conversion testing, relative chromosomal location comparison and estimated time for gene duplication, we believe that rodent PL-I and PL-II genes are species-specific and are the results of serial duplications which occurred after the divergence of mouse and rat. Our analysis also reveals that continual gene duplication and divergence occurred during the evolution of rodent PRL gene family.     

Rapid expansion of the Ly49 gene cluster in rat

The cytotoxic activity of mouse natural killer cells is regulated in part through cell surface molecules belonging to the Ly49 multigene family. In mice, the genomic sequence of the Ly49 gene cluster has been examined in detail and this analysis provided a model of the expansion of this multigene family. In the present study, we have analyzed a 1.8-Mb region of the draft rat genome revealing surprising differences in size and gene content between the mouse and the rat Ly49 clusters. The rat cluster contains at least 36 Ly49 genes, including pseudogenes, while dot-plot analysis of the cluster reveals an equidistant spacing of genes, suggesting that duplication of genes in the cluster occurred through a mechanism similar to that in the mouse. Phylogenetic analysis of the predicted rat genes reveals a number of distinct gene clusters and indicates that the majority of gene duplication events occurred after the divergence of mice and rats. Thus, the rodent Ly49 locus is subject to extremely rapid gene amplification and diversification.     

Species-specific size expansion and molecular evolution of the oleosins in angiosperms

Oleosins are hydrophobic plant proteins thought to be important for the formation of oil bodies, which supply energy for seed germination and subsequent seedling growth. To better understand the evolutionary history and diversity of the oleosin gene family in plants, especially angiosperms, we systematically investigated the molecular evolution of this family using eight representative angiosperm species. A total of 73 oleosin members were identified, with six members in each of four monocot species and a greater but variable number in the four eudicots. A phylogenetic analysis revealed that the angiosperm oleosin genes belonged to three monophyletic lineages. Species-specific gene duplications, caused mainly by segmental duplication, led to the great expansion of oleosin genes and occurred frequently in eudicots after the monocot–eudicot divergence. Functional divergence analyses indicate that significant amino acid site-specific selective constraints acted on the different clades of oleosins. Adaptive evolution analyses demonstrate that oleosin genes were subject to strong purifying selection after their species-specific duplications and that rapid evolution occurred with a high degree of evolutionary dynamics in the pollen-specific oleosin genes. In conclusion, this study serves as a foundation for genome-wide analyses of the oleosins. These findings provide insight into the function and evolution of this gene family in angiosperms and pave the way for studies in other plants.     

Phylogenetic Analysis of the Cytochrome P450 3 (CYP3) Gene Family

Cytochrome P450 genes (CYP) constitute a superfamily with members known from the Bacteria, Archaea, and Eukarya. The CYP3 gene family includes the CYP3A and CYP3B subfamilies. Members of the CYP3A subfamily represent the dominant CYP forms expressed in the digestive and respiratory tracts of vertebrates. The CYP3A enzymes metabolize a wide variety of chemically diverse lipophilic organic compounds. To understand vertebrate CYP3 diversity better, we determined the killifish (Fundulus heteroclitus) CYP3A30 and CYP3A56 and the ball python (Python regius) CYP3A42 sequences. We performed phylogenetic analyses of 45 vertebrate CYP3 amino acid sequences using a Bayesian approach. Our analyses indicate that teleost, diapsid, and mammalian CYP3A genes have undergone independent diversification and that the ancestral vertebrate genome contained a single CYP3A gene. Most CYP3A diversity is the product of recent gene duplication events. There is strong support for placement of the guinea pig CYP3A genes within the rodent CYP3A diversification. The rat, mouse, and hamster CYP3A genes are mixed among several rodent CYP3A subclades, indicative of a complex history involving speciation and gene duplication. Phylogenetic analyses suggest two CYP3A gene duplication events early in rodent history, with the rat CYP3A9 and mouse Cyp3a13 clade having a sister relationship to all other rodent CYP3A genes. In primate history, the human CYP3A43 gene appears to have a sister relationship to all other known primate CYP3A genes. Other, more recent gene duplications are hypothesized to have occurred independently within the human, pig, rat, mouse, guinea pig, and fish genomes. Functional analyses suggest that gene duplication is strongly tied to acquisition of new function and that convergent evolution of CYP3A function may be frequent among independent gene copies. Current address (Rachel L. Cox): Laboratory of Aquatic Biomedicine, Marine Biology Laboratory, Woods Hole, MA 02543, USA     

Analysis of the evolution of angiotensin II type 1 receptor gene in mammals (mouse, rat, bovine and human).

The nucleotide and amino acid sequences for mouse angiotensin II (AII) type 1A and 1B receptors were deduced from their complementary and genomic DNAs. Evolutionary analyses based on the nucleotide sequences of the coding region of AII type 1 receptor genes indicated that the duplication event of the type 1 gene occurred 24 +/- 2 million years ago before the divergence between the rat and mouse but after the divergence between rodents and the human/artiodactyls couple. This conclusion was consistent with the results of genomic Southern blot analyses, which revealed that the mouse and rat possess 2 similar but separate genes, whereas the bovine and human have only a single class gene.     

Comparable Rates of Gene Loss and Functional Divergence after Genome Duplications Early in Vertebrate Evolution

Duplicated genes are an important source of new protein functions and novel developmental and physiological pathways. Whereas most models for fate of duplicated genes show that they tend to be rapidly lost, models for pathway evolution suggest that many duplicated genes rapidly acquire novel functions. Little empirical evidence is available, however, for the relative rates of gene loss vs. divergence to help resolve these contradictory expectations. Gene families resulting from genome duplications provide an opportunity to address this apparent contradiction. With genome duplication, the number of duplicated genes in a gene family is at most 2(n), where n is the number of duplications. The size of each gene family, e.g., 1, 2, 3, . . . , 2(n), reflects the patterns of gene loss vs. functional divergence after duplication. We focused on gene families in humans and mice that arose from genome duplications in early vertebrate evolution and we analyzed the frequency distribution of gene family size, i.e., the number of families with two, three or four members. All the models that we evaluated showed that duplicated genes are almost as likely to acquire a new and essential function as to be lost through acquisition of mutations that compromise protein function. An explanation for the unexpectedly high rate of functional divergence is that duplication allows genes to accumulate more neutral than disadvantageous mutations, thereby providing more opportunities to acquire diversified functions and pathways.     

Localization of placental lactogen-I in trophoblast giant cells of the mouse placenta

The purpose of this investigation was to identify the cellular origin of placental lactogen-I (PL-I) expression in the mouse placenta and to cytologically define the transition from PL-I to PL-II expression during gestation. PL-I mRNA expression was assessed by in situ hybridization, and expression of PL-I and PL-II protein was determined by immunocytochemical analysis. PL-I mRNA and protein were localized to trophoblast giant cells. Trophoblast giant cells ceased producing PL-I at midgestation and began expressing PL-II. PL-I immunoreactivity was present in trophoblast giant cells on Days 9 and 10 of gestation but was not detectable in trophoblast giant cells on Day 11 of gestation. Immunoreactive PL-II-producing giant cells were detected first on Day 10 of gestation, continuing on Day 11 of gestation. Expression of PL-I and PL-II signals a significant functional transition in trophoblast giant cells of the developing mouse placenta.     

Rapid lineage-specific diversification of the mast cell chymase locus during mammalian evolution

Serine proteases constitute the major protein granule content of cells of several hematopoietic cell lineages. A subgroup of these proteases, including the mast cell chymases, neutrophil cathepsin G, and T cell granzymes B to F and N, are in all investigated mammals encoded in one locus, the chymase locus. It is interesting to note that this locus has diversified greatly during the last 95 Myr of mammalian evolution. This divergence is exemplified by the presence of Mcpt8-related genes and multiple β-chymases in the mouse and rat, which lack direct counterparts in primates and in seven functional granzyme genes in the mouse where the human locus has only two. To study the expansion of the locus during rodent evolution and to better understand the evolutionary origin of β-chymases and the Mcpt8-family, we have performed a detailed analysis of the chymase locus of four mammalian species, i.e., human, dog, mouse, and rat. As a result, we report here a second chymase-like gene in dog, Cma2, which clusters with β-chymases in phylogenetic analyses. This finding supports a duplication of the common ancestor for α- and β-chymases before the major radiation of placental mammals, and a loss of the ancestral β-chymase gene sometime during primate evolution. Moreover, we show that in the rat, the Mcpt8-family diversified relatively recently together with sequences related to the β-chymase Mcpt2. Eight novel genes were identified in the duplication region, four of which are predicted to be functional. Duplications of rat granzyme B- and C-like sequences occurred seemingly independently within a similar time frame, but did not give rise to functional genes. Due to the duplications in rat and deletions in the carnivore/primate lineage, the rat chymase locus is approximately 15 and 9 times larger than its counterparts in dog and human, respectively. These findings illustrate the importance of gene duplications in conferring rapid changes in mammalian genomes.     

The human and murine protocadherin-beta one-exon gene families show high evolutionary conservation, despite the difference in gene number

Extensive cDNA analysis demonstrated that all human and mouse protocadherin-beta genes are one-exon genes. The protein sequences of these genes are highly conserved, especially the three most membrane-proximal extracellular domains. Phylogenetic analysis suggested that this unique gene family evolved by duplication of one single protocadherin-beta gene to 15 copies. The final difference in the number of protocadherin-beta genes in man (#19) and mouse (#22) is probably caused by duplications later in evolution. The complex relationship between human and mouse genes and the lack of pseudogenes in the mouse protocadherin-beta gene cluster suggest a species-specific evolutionary pressure for maintenance of numerous protocadherin-beta genes.