Prediction of sites under adaptive evolution in flavin-containing monooxygenases: Selection pattern revisited

Flavin-containing monooxygenase (FMO), like cytochrome P450 (CYP), is a monooxygenase that uses the reducing equivalents of NADPH to reduce one atom of molecular oxygen to water, while the other atom is used to oxidize the substrate. Recently, it was shown that some CYP isoforms have been subject to positive selection. However, it is unknown whether the highly conserved phase I detoxification enzyme, FMO, has undergone similar positive Darwinian selection. We used maximum-likelihood models of codon substitution, evolutionary fingerprinting, and cross species comparison to investigate the occurrence of adaptive evolution in FMO sequences. We used recent genomic data from a range of species, including vertebrates and invertebrates. We present the evidence for the occurrence of adaptive evolution in mammalian FMO 3, 4, 5, and fugu FMOs but not in mammalian FMO 1, FMO 2, frog FMOs, other fish FMOs and invertebrate FMOs. The sites under adaptive evolution were significantly associated with the insertion domain in mammalian FMO 5. We identified specific amino acid sites in FMOs 3–5 that are likely targets for selection based on the patterns of parallel amino acid change. The most likely role of adaptive evolution is the repair of mutations that permitted optimal NADP+ binding and improved catalytic efficiency. The occurrence of positive selection during the evolution of phase I detoxification enzymes such as FMOs 3–5 and fugu FMO suggests the occurrence of both high selection pressure acting on species within their unique habitats and significant changes in intensity and direction (forms of xenobiotics and drugs) resulting from changes in microhabitat and food.     

Episodic evolution of prolactin gene in primates

In the present study, we obtained exon 2--5 of prolactin (PRL) gene from four primate species by PCR and sequencing. Adding other genes available in GenBank, we calculate amino acid substitution rates for prolactin gene in primate. Comparison of nonsynonymous substitution rate to synonymous substitution rate ratios shows no evidence of positive selection for any lineage of primate prolactin gene.According to this and the facts that (i) no sites under positive selection are inferred by using maximum-likelihood method;(ii) among 32 amino acid replacement that occurred along the rapid evolutionary phase, only two are included in the 40 functionally important residues, indicating that amino acid replacement tends to occur in those functionally unimportant residues; (iii) partial of prolactin function is replaced by placental lactogen in primate at the rapid evolutionary phase of prolactin gene, we thus deem that it is relaxation of purifying selection to some extent rather than positive selection that enforces the rapid evolution of primate prolactin gene.     

水龙骨科附生蕨类Rubisco大亚基的适应性进化:正向选择位点的鉴定

 核酮糖-1,5-二磷酸羧化酶/加氧酶（Rubisco,EC 4.1.1.39）在植物光合作用中起重要作用,既负责碳同化又引发光呼吸。催化固定CO₂的活性位点位于Rubisco大亚基,由叶绿体rbc L基因编码。水龙骨科是现存蕨类中最为衍生的类群,多为附生植物。为验证蕨类植物在白垩纪适应被子植物兴起而发生分化的观点,本研究以水龙骨科附生蕨类为对象,利用位点间可变ω（非同义替换率d_N和同义替换率d_S的比值）模型对其rbc L基因的适应性进化进行分析。通过比较模型M1a/M2a 和模型M7/M8,在氨基酸水平上,共鉴定出7个正向选择位点：133L,251M,262A,265V,282Y,326I和362I。其中位点262A,265V,282Y及326I对维持Rubisco功能的重要性也得到已发表实验数据的支持。这些结果一方面显示了基于ω比值检验DNA编码序列分子适应的有效性,另一方面也提示水龙骨类可借助rbcL等功能基因的适应性进化,应对白垩纪被子植物引发的陆地生态系统改变。     

Molecular evolution of connective tissue growth factor in Cyprinidae (Teleostei: Cypriniformes)

Connective tissue growth factor (CTGF) plays an important role in regulation of cell growth, differentiation, apoptosis and individual development in animals. The study of sequences variation and molecular evolution of CTGF gene across various species of the cyprinid could be helpful for understanding of speciation and gene divergence in this kind of fish. In this study, 19 novel sequences of CTGF gene were obtained from the representative species of the family Cyprinidae using PCR amplification, cloning and sequencing. Phylogenetic relationships of Cyprinidae were reconstructed by neighbor-joining (NJ), maximum parsimony (MP), maximum likelihood (ML) and Bayesian method. Oryzias latipes from the family Cyprinodontidae was assigned to be the outgroup taxon. Leuciscini and Barbini were clustered into the monophyletic lineages respectively with the high nodal supports. The estimation of the ratio of nonsynonymous to synonymous substitution (dN/dS) for the various branches indicated that there stood the different evolution rates between the Leuciscini and the Barbini. With the ratio of dN/dS of the Leuciscini being lower than that of the Barbini, species within the Barbini were demonstrated to be subjected to the relatively less selection pressure and under the relaxable evolution background. A 6 bp indel (insertion/deletion) was found at the 5' end of CTGF gene of Cyprinidae, and this 6 bp deletion only appeared in the Leuciscini, which is a typical characteristic of the Leuciscini and provides evidence for the monophylogeny of the Leuciscini. For the amino acid sequences of CTGF protein, the most variations and Indels were distributed in the signal region and IGFBP region of this protein, implying that these variations were correlated with the regulation of the CTGF gene expression and protein activity.     

Molecular evolution of connective tissue growth factor in Cyprinidae （Teleostei： Cypriniformes）

Connective tissue growth factor （CTGF） plays an important role in regulation of cell growth, differentiation, apoptosis and individual development in animals. The study of sequences variation and molecular evolution of CTGF gene across various species of the cyprinid could be helpful for understanding of speciation and gene divergence in this kind of fish. In this study, 19 novel sequences of CTGF gene were obtained from the representative species of the family Cyprinidae using PCR amplification, cloning and sequencing. Phylogenetic relationships of Cyprinidae were reconstructed by neighbor-joining （NJ）, maximum parsimony （MP）, maximum likelihood （ML） and Bayesian method. Oryzias latipes from the family Cyprinodontidae was assigned to be the outgroup taxon. Leuciscini and Barbini were clustered into the monophyletic lineages, respectively, with the high nodal supports. The estimation of the ratio of non-synonymous to synonymous substitution （dN/dS） for the various branches indicated that there stood the different evolution rates between the Leuciscini and the Barbini. With the ratio of dN/dS of the Leuciscini being lower than that of the Barbini, species within the Barbini were demonstrated to be subjected to the relatively less selection pressure and under the relaxable evolution background. A 6 bp indel （insertion/deletion） was found at the 5＇ end of CTGF gene of Cyprinidae, and this 6 bp deletion only appeared in the Leuciscini, which is a typical characteristic of the Leuciscini and provides evidence for the monophylogeny of the Leuciscini. For the amino acid sequences of CTGF protein, the most variations and indels were distributed in the signal region and IGFBP region of this protein, implying that these variations were correlated with the regulation of the CTGF gene expression and protein activity.     

Adaptive evolution of non-coding DNA in Drosophila

A large fraction of eukaryotic genomes consists of DNA that is not translated into protein sequence, and little is known about its functional significance. Here I show that several classes of non-coding DNA in Drosophila are evolving considerably slower than synonymous sites, and yet show an excess of between-species divergence relative to polymorphism when compared with synonymous sites. The former is a hallmark of selective constraint, but the latter is a signature of adaptive evolution, resembling general patterns of protein evolution in Drosophila. I estimate that about 40-70% of nucleotides in intergenic regions, untranslated portions of mature mRNAs (UTRs) and most intronic DNA are evolutionarily constrained relative to synonymous sites. However, I also use an extension to the McDonald-Kreitman test to show that a substantial fraction of the nucleotide divergence in these regions was driven to fixation by positive selection (about 20% for most intronic and intergenic DNA, and 60% for UTRs). On the basis of these observations, I suggest that a large fraction of the non-translated genome is functionally important and subject to both purifying selection and adaptive evolution. These results imply that, although positive selection is clearly an important facet of protein evolution, adaptive changes to non-coding DNA might have been considerably more common in the evolution of D. melanogaster.     

Evidence for host-cell selection of influenza virus antigenic variants

Extensive antigenic variability and a capricious epidemiology are characteristics of influenza A and B viruses of man. The haemagglutinin (HA) undergoes frequent and progressive antigenic drift as a result of selection, under immunological pressure, of viruses possessing alterations in the amino acid sequences at specific sites in the molecule. Here we present evidence for an additional selection mechanism for antigenic variants of influenza virus that depends on differing host cell tropisms of virus subpopulations. These studies were initiated after earlier observations of the occurrence of a marked degree of antigenic variation during passage of laboratory strains of influenza virus in eggs and cell cultures (J.C.J., in preparation). We have now shown that cultivation of influenza B viruses in eggs selects subpopulations which are antigenically distinct from virus from the same source grown in mammalian cell cultures. As antigenic characterization of influenza virus strains for epidemiological purposes and for the preparation of influenza vaccines conventionally relies on the cultivation of virus in eggs, our findings may have important practical implications for vaccine design and efficacy.     

Selection pressure drives the co-evolution of several lipid metabolism genes in mammals

Adipose tissue is an important endocrine organ and energy supplier.Its physiological effect on the regulation of the energy balance is considered an important factor underlying the evolution of mammals.To test whether the genes controlling lipid metabolism have undergone adaptive molecular change in the evolution of mammals,in this study,we used the orthologous gene sequences of 12 important lipid metabolism proteins (leptin,OB-RL,RXRA,RXRB,RXRG,PPARA,PPARB/D,PPARG,PNLIP,ADIPOQ,LPL and UCP1) from NCBI’s databases.We found evidence that 4 of the corresponding genes (leptin,ADIPOQ,PNLIP and PPARA) have undergone positive selection in their evolutionary history and that most adaptive changes occurred during the evolution of the super-clades Laurasiatheria (placentals) and suborders within Euarchontoglires (primates and rodents).Comparisons across sets of genes showed that in a third of cases,bursts of positive selection,more than would be expected by chance,occurred on corresponding branches.We propose that the positive selection drives adaptive changes in some lipid metabolism genes in or within Laurasiatheria and Euarchontoglires clades.Along with evidence from earlier studies,our results show that co-evolution among interacting lipid metabolism proteins has taken place.     

分子序列的突变危险性与遗传密码的编码规则

定义了分子序列（碱基序更和对应的氨基酸序列）的突变危险性函数，通过危险性函数的极小化可以导出遗传密码的简并规则，终止密码子在密码表上的位置及亲－水畴排布，氨基酸在密码表上的定位接近标准密码表，各个生物类别的突变危险性函数值差别不超过５％，可看作守恒量，Ｒａｓ家庭癌基因产物具有较大的突变危险性函数值，这提示分子序列的突变危险性函数可能有重要的应用价值和理论意义。     

A complete sequence and comparative analysis of a SARS-associated virus(Isolate BJ01)

Severe Acute Respiratory Syndrome (SARS) is a newly identified infectious disease[1—5]. The global outbreak of SARS has been threatening the health of people worldwide and has killed 353 people and infected more than 5462 in 27 countries, as reported by WHO on April 29, 2003 (http://www.who.int/csr/sarscountry/en). Although it has been recognized that a variant of virus from the family of coronavirus might be the candidate pathogen of SARS[1—5], its identity as the unique pathogen sti…     

A complete sequence and comparative analysis of a SARS-associated virus （Isolate BJO1）

The genome sequence of the Severe Acute Respiratory Syndrome (SARS)-assoclated virus provides essential information for the identification of pathogen(s), exploration of etiology and evolution, interpretation of transmission and pathogenesis, development of diagnostics, prevention by future vaccination, and treatment by developing new drugs.We report the complete genome sequence and comparative analysis of an isolate (B J01) of the coronavirus that has been recognized as a pathogen for SARS. The genome is 29725 nt in size and has 11 ORFs (Open Reading Frames). It is composed of a stable region encoding an RNA-dependent RNA polymerase (composed of 20RFs) and a variable region representing 4 CDSs (coding sequences) for viral structural genes (the S, E, M, N proteins) and 5 PUPs (putative uncharacterized proteins). Its gene order is identical to that of other known coronaviruses. The sequence alignment with all known RNA viruses places this virus as a member in the family of Coronaviridae. Thirty putative substitutions have been identified by comparative analysis of the 5 SARS-associated virus genome sequences in GenBank. Fifteen of them lead to possible amino acid changes (non-synonymous mutations) in the proteins. Three amino acid changes, with predicted alteration of physical and chemical features, have been detected in the S protein that is postulated to be involved in the immunoreactions between the virus and its host.Two amino acid changes have been detected in the M protein,which could be related to viral envelope formation. Phylogenetic analysis suggests the possibility of non-human origin of the SARS-associated viruses but provides no evidence that they are man-made. Further efforts should focus on identifying the etiology of the SARS-associated virus and ruling out conclusively the existence of other possible SARS-related pathogen(s).     

禾本科植物β-淀粉酶基因家族分子进化及响应非生物胁迫的表达模式分析

 β-淀粉酶(beta-amylase,BAM)是一类关键的淀粉水解酶,在禾谷类作物生长发育过程中起着重要作用,与植物多种非生物胁迫响应相关.本研究通过系统发育分析,将水稻、玉米、高粱、谷子、二穗短柄草5 种禾本科植物中共54 个BAM 基因分为10 个同源基因簇,每个同源基因簇都涵盖了这5 个物种,因此推测在禾本科祖先物种中至少含有10 个BAM 基因,并且在禾本科植物分化后没有发生明显的基因丢失事件.基于对编码蛋白质序列的功能分化分析,表明同源基因簇间存在明显的进化速率的差异.对10 个同源基因簇进行了适应性进化检测,发现有3 个同源簇在禾本科植物的进化过程中经历了适应性进化.此外,对水稻β-淀粉酶的表达分析发现,一些β-淀粉酶具有组织特异性表达特征,并且至少有5 个水稻的β-淀粉酶基因具有受到非生物逆境的胁迫而表现出不同的表达模式.本研究结果为进一步探讨禾本科BAM 基因的生物学功能提供了一定的理论基础.     

Vertical adaptive radiation in ocean Prochlorococcus: Evolutionary implications of the Chl b/a ratio from molecular evidence

Prochlorococcus is a marine cyanobacterium of global significance. Two ecotypes are adapted to either high-light (HL) or low-light (LL) conditions. The ratio between chlorophyll (Chl) a and b is a distinguishing characteristic of these two ecotypes. However, how this ratio evolved in Prochlorococcus during this ecotype differentiation remains unclear. Our analyses reveal that the ancestor of Prochlorococcus was typically low-light adapted. The LL ecotype showed a stagnant evolution, and the HL ecotype was recently diverged. There was an adaptive radiation after directional evolution in the Chl b/a ratio regulation. Recombination in chlorophyllide a oxygenase (CAO) and positive selection on Clp protease contributed to the directional evolution of Prochlorococcus. The recombinant fragments of CAO were correlated with a large group of shared coevolving sites. Evidence of positive selection was found in both subunits of Clp. Chl b/a ratio evolution, as annotated by molecular evidence, appears to be among the crucial reasons that explain how Prochlorococcus has become the dominant photosynthetic organism in the ocean.     

基于叶绿体基因组全序列分析真叶植物叶绿体基因的适应性进化

 真叶植物包括蕨类、裸子植物和被子植物。迄今已积累有较为丰富的真叶植物叶绿体基因组全序列数据。选取了29种真叶植物的叶绿体基因组全序列,采用PAML 软件基于位点间可变ω模型,分别分析了蕨类、裸子植物和被子植物叶绿体基因的适应性进化。结果显示：① 蕨类、裸子植物和被子植物各有6.5%、7.5%和19.2%的叶绿体基因受正选择作用;被子植物经历正选择的叶绿体基因明显比蕨类和裸子植物为多;② 被正选择作用的叶绿体基因主要是遗传系统和光合系统基因,它们的编码产物涉及叶绿体蛋白质合成、基因转录、能量转化与调节及光合作用等过程。推测叶绿体功能基因可能在真叶植物对陆生生态环境的适应过程中起着重要作用。     

Genome organization and phylogenetic tree analysis of Garlic virus E, a new member of genus Allexivirus

The complete sequence of an Allexivirus isolated from garlic plants in Yuhang City, Zhejiang Province, China had been determined. The single-strand, positive RNA genome was 8451 nucleotides in length excluding poly(A) tail. The genome organization of this virus was similar to that of the other Allexiviruses but only with 62.8%-64.8% nucleotide acid identities. The amino acid sequences of proteins encoded by ORF1-6 shared 67.6%-78.5%, 55.4%-66.2%, 56.7%- 66.4%, 40.3%-55.6%, 66.3%-79.7% and 52.2%- 68.8% identities with those of the others respectively. The homology range between it and the other Allexiviruses was similar to that between the other distinct species in this genus. A more comprehensive comparison using all available CP amino acid sequences showed that it shared only 63.9%- 79.8% amino acids identical with the others. Therefore, it had been considered as a new member of the genus, named as garlic virus E (GarV-E). Phylogenetic analysis confirmed GarV-E as a distinct member and the correct names and classification of some members of genus Allexivirus were also discussed.     

Genetic insight of the H5N1 hemagglutinin cleavage site

The cleavability of the hemagglutinin (HA) plays a major role in virulence of avian influenza viruses. Detailed analyses of the cleavage sequences and their evolution would give insights into the high pathogenicity of the H5N1 virus. HA segments were visually identifiable in the cellular automata (CA) image, and a feature gene segment (FGS) was only found in H5N1 rather than any other subtype. This FGS is a 30-bp gene segment mainly consisting of ‘A’ and ‘G’. When translated into amino acids the FGS converted into a sequence of mainly basic amino acids with positive charges. This feature amino acid segment (FAAS) was located in the cleavage site loop of HA which was potentially cleavable by various proteases. The 3D structure of H5N1 HA was reconstructed using homology modelling. It was found that the cleavage site loop was well exposed to potential proteases. The molecular surfaces were reconstructed to study how mutation and deletion of some amino acids in the FAAS affected the charge distribution. It was found that some mutations had severely changed the landscape of the charge dis- tribution. Statistical analyses of FAAS were made with respect to when and where the H5N1 viruses were found. In 2005, there were less un-mutated FAAS than the other years according to temporal evolution, and more mutated FAAS appeared in China than other regions according to geographic dis- tribution. These results are helpful for exploring the evolution of virus high pathogenicity.     

基于Kalman滤波的自适应MIMO信道估计

给出了一种基于Kalman滤波的自适应MIMO信道估计方法，分析了算法的剩余误差能量随迭代次数增加的变化情况，对饱和MIMO系统和欠饱和MIMO系统，分别采用Kalman自适应MIMO信道估计方法和最大似然信道估计方法，进行了数值仿真和对比。从对比的结果来看，文中给出的Kalman自适应信道估计具有和最大似然信道估计相近的性能，而在饱和系统中，Kalman估计算法性能要略好一些。     

Phylogenetic origins and adaptive evolution of avian and mammalian haemoglobin genes

Recent years have seen rapid growth in amino acid sequence data on globins and nucleotide sequence data on haemoglobin genes and pseudogenes, and cladistic analysis of these data continues to reveal new facets of globin evolution. Our present findings demonstrate: (1) avian and mammalian embryonic alpha genes (pi and xi, respectively) had a monophyletic origin involving an alpha locus duplication about 400 Myr ago soon after the duplication which separated alpha and beta genes; (2) much later in phylogeny, independent beta-gene duplications produced the embryonic rho locus of birds and embryonic epsilon and fetal gamma loci of mammals. This parallels the earlier finding that myoglobins evolved more than once from generalized globin ancestors. Here we support the view that such globin evolution resulted from natural selection acting on mutations in duplicated genes. Thus, our evidence contradicts the neutralist view in which almost all amino acid substitutions in descent to extant globins evaded positive selection.     

Escherichia coli K-12 undergoes adaptive evolution to achieve in silico predicted optimal growth

Annotated genome sequences can be used to reconstruct whole-cell metabolic networks. These metabolic networks can be modelled and analysed (computed) to study complex biological functions. In particular, constraints-based in silico models have been used to calculate optimal growth rates on common carbon substrates, and the results were found to be consistent with experimental data under many but not all conditions. Optimal biological functions are acquired through an evolutionary process. Thus, incorrect predictions of in silico models based on optimal performance criteria may be due to incomplete adaptive evolution under the conditions examined. Escherichia coli K-12 MG1655 grows sub-optimally on glycerol as the sole carbon source. Here we show that when placed under growth selection pressure, the growth rate of E. coli on glycerol reproducibly evolved over 40 days, or about 700 generations, from a sub-optimal value to the optimal growth rate predicted from a whole-cell in silico model. These results open the possibility of using adaptive evolution of entire metabolic networks to realize metabolic states that have been determined a priori based on in silico analysis.