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The Human Papillomavirus Database.

Papillomaviruses are responsible for a variety of diseases in humans and animals, ranging from harmless skin warts to lethal cancers. They also make up one of the most genetically diversified families of viruses known, and could represent a model system of DNA-virus evolution. A specialized genetic sequences database, The Human Papillomavirus Database and Analysis Project, was recently established in an effort to provide database services that are specific to papillomaviruses to the research community and to perform a variety of sequence-based analyses. This review is intended to present the scope of the information currently contained in the database and to outline some of the analyses that have been performed on the genetic sequences. These analyses will address issues including phylogenetic relationships, recombination events, selective pressures on different genes and the possibility of cross-species transmission in the case of the papillomaviruses. Copyright 1995 S. Karger AG, Basel

Journal Article↗

Geographical prevalence of two types of Epstein-Barr virus.

The Jijoye EBV strain is characterized by a substitution of 1.8 kb in the C-terminal part of the EBNA 2 gene compared to B95-8 or M-ABA virus. This made it possible to construct hybridization probes specific for M-ABA (type A) and Jijoye viruses (type B), which have been used to type the EBV genomes in 38 spontaneously established cell lines. Type A is more prevalent being found in 31 of 38 cases; type B virus was found in five cell lines (Jijoye, LY 67, QIMR-GOR, BL 16, and BL 29); and two cell lines, Daudi and EB-3, contained neither the M-ABA- nor the Jijoye-specific sequences. EBV type B appears to be less ubiquitous, since all type B isolates, including AG 876 virus, originated from Central Africa, La Réunion, and New Guinea. All the other cell lines, carrying EBV type A, were established from patients from Central Africa (4), North Africa (7), New Guinea (1), and Asia (6) and from white individuals (13). The restricted geographical localization of EBV type B in parts of the southern hemisphere and its similarity to herpesvirus papio (T. Dambaugh, K. Hennessy, L. Chamnankit, and E. Kieff (1984) Proc. Natl. Acad. Sci. USA 81, 7632-7636) could suggest that such viruses may have evolved by recombination of EBV with a related Old World monkey virus, alternatively, evolution of virus variants within the human species also being conceivable.

Cloning, Molecular↗

A porcine rotavirus strain with dual VP7 serotype specificity.

Porcine rotavirus MDR-13, which on original isolation showed a two-way antigenic relationship with human rotavirus RV-3, shows VP7 relationships with serotype G5 as well as G3 viruses upon gene reassortment. Analysis of porcine MDR-13 and the MD-UK reassortant revealed marked nucleotide and amino acid similarity of VP7 genes of these viruses with those of both serotype G3 and G5 viruses. Evolution of such a strain, possibly by sequential mutations in the VP7 gene, is discussed.

Amino Acid Sequence↗

RNA genome stability of Toscana virus during serial transovarial transmission in the sandfly Phlebotomus perniciosus.

We have carried out a T1 ribonuclease fingerprinting analysis of the RNA genomes of Toscana virus isolates from successive generations of an experimentally virus-infected laboratory colony of Phlebotomus perniciosus sandflies. This analysis detected no virus RNA genome changes during transovarial transmission of the virus over 12 sandfly generations (a period of almost 2 years). These results demonstrate that although RNA viruses can exhibit high rates of mutational change under a variety of conditions, Toscana virus RNA genomes can be maintained in a stable manner during repeated transovarial virus transmission in the natural insect host. The implications of these results for insect RNA virus evolution are discussed.

Animals↗

A real-time RT-PCR assay for quantifying the fitness of tobacco etch virus in competition experiments.

Relative fitness determination has become a standard tool in experimental virus evolution studies. In this type of studies, the tested strain is mixed with a reference strain, which differs in an easy-to-score and genetically stable marker, and allowed to compete for a limited common pool of resources during a given number of generations. In this report, a TaqMan real-time PCR methodology is proposed for quantifying the relative fitness of tobacco etch potyvirus strains (TEV) in in planta mixed infections with a reference TEV strain. Two different forward primers along with a common reverse one are used into separated reactions mixes from the same RNA preparation. The reference strain, named TEV-PC1, was genetically engineered to carry a neutral marker in a highly conserved region of the RNA polymerase NIb gene. This marker allows tracking the frequency of both competitors during competition experiments by real-time quantitative PCR using specific primers. Both the reproducibility and sensitivity of the method have been explored. Reproducibility was assessed by running multiple competition experiments for the same genotype. Sensitivity was assessed by comparing the results of competition experiments against TEV-PC1 of 24 single-nucleotide substitutions mutants.

Gene Amplification↗

A map of the diversity of RNA3 recombinants appearing in plants infected with Cucumber mosaic virus and Tomato aspermy virus.

In order to better understand the role of recombination in creating the diversity of viral genomes that is acted on by selection, we have studied in detail the population of recombinant RNA3 molecules occurring in tobacco plants coinfected with wild-type strains of cucumber mosaic virus (CMV) and tomato aspermy virus (TAV) under conditions of minimal selection pressure. Recombinant RNA3s were observed in 9.6% of the samples. Precise homologous recombination predominated since it was observed at 28 different sites, primarily in six hot spots. Imprecise homologous recombination was observed at two sites, particularly within a GU repeat in the 5' noncoding region. Seven of the eight aberrant homologous recombination sites observed were clustered in the 3' noncoding region. These results have implications on the role of recombination in host adaptation and virus evolution. They also provide essential baseline information for understanding the potential epidemiological impact of recombination in transgenic plants expressing viral sequences.

Base Sequence↗

The immunogenicity of human and murine cytomegaloviruses.

Cytomegaloviruses are strictly host-species-specific. During an aeon of co-evolution, virus and host have found an arrangement: the productive and cytopathogenic cycle of viral gene expression is held in check by the host's immune response. As a consequence, cytomegalovirus disease is restricted to the immunocompromised host. The virus has evolved strategies to avoid its elimination and eventually hides itself in a silent state, referred to as 'viral latency'. Redundant molecular mechanisms have been identified by which cytomegaloviruses interfere with antigen presentation pathways to 'evade' immune control. In the annual period covered by this review, the IE1 protein was revisited as an immunodominant antigen of human cytomegalovirus and the identification of a first antigenic early-phase peptide of murine cytomegalovirus that escapes viral immunosubversive mechanisms may initiate a period of research on the immune control of cytomegaloviruses 'beyond immune evasion'.

Animals↗

De novo discovery of conserved gene clusters in microbial genomes with Spacedust.

Metagenomics has revolutionized environmental and human-associated microbiome studies. However, the limited fraction of proteins with known biological processes and molecular functions presents a major bottleneck. In prokaryotes and viruses, evolution favors keeping genes participating in the same biological processes colocalized as conserved gene clusters. Conversely, conservation of gene neighborhood indicates functional association. Here we present Spacedust, a tool for systematic, de novo discovery of conserved gene clusters. To find homologous protein matches, Spacedust uses fast and sensitive structure comparison with Foldseek. Partially conserved clusters are detected using novel clustering and order conservation P values. We demonstrate Spacedust's sensitivity with an all-versus-all analysis of 1,308 bacterial genomes, identifying 72,843 conserved gene clusters containing 58% of the 4.2 million genes. It recovered 95% of antiviral defense system clusters annotated by the specialized tool PADLOC. Spacedust's high sensitivity and speed will facilitate the annotation of large numbers of sequenced bacterial, archaeal and viral genomes.

Metagenomics↗

Complete alanine scanning of intersubunit interfaces in a foot-and-mouth disease virus capsid reveals critical contributions of many side chains to particle stability and viral function.

Spherical virus capsids are large, multimeric protein shells whose assembly and stability depend on the establishment of multiple non-covalent interactions between many polypeptide subunits. In a foot-and-mouth disease virus capsid, 42 amino acid side chains per protomer are involved in noncovalent interactions between pentameric subunits that function as assembly/disassembly intermediates. We have individually truncated to alanine these 42 side chains and assessed their relevance for completion of the virus life cycle and capsid stability. Most mutations provoked a drastic reduction in virus yields. Nearly all of these critical mutations led to virions whose thermal inactivation rates differed from that of the parent virus, and many affected also early steps in the viral cycle. Rapid selection of genotypic revertants or variants with forward or compensatory mutations that restored viability was occasionally detected. The results with this model virus indicate the following. (i). Most of the residues at the interfaces between capsid subunits are critically important for viral function, in part but not exclusively because of their involvement in intersubunit recognition. Each hydrogen bond and salt bridge buried at the subunit interfaces may be important for capsid stability. (ii). New mutations able to restore viability may arise frequently at the subunit interfaces during virus evolution. (iii). A few interfacial side chains are functionally tolerant to truncation and may provide adequate mutation sites for the engineering of a thermostable capsid, potentially useful as an improved vaccine.

Alanine↗

A patient with asymptomatic severe acute respiratory syndrome (SARS) and antigenemia from the 2003-2004 community outbreak of SARS in Guangzhou, China.

An asymptomatic case of severe acute respiratory syndrome (SARS) occurred early in 2004, during a community outbreak of SARS in Guangzhou, China. This was the first time that a case of asymptomatic SARS was noted in an individual with antigenemia and seroconversion. The asymptomatic case patient and the second index case patient with SARS in the 2003-2004 outbreak both worked in the same restaurant, where they served palm civets, which were found to carry SARS-associated coronaviruses. Epidemiological information and laboratory findings suggested that the findings for the patient with asymptomatic infection, together with the findings from previously reported serological analyses of handlers of wild animals and the 4 index case patients from the 2004 community outbreak, reflected a likely intermediate phase of animal-to-human transmission of infection, rather than a case of human-to-human transmission. This intermediate phase may be a critical stage for virus evolution and disease prevention.

Antigens, Viral↗

Lamivudine-resistant human immunodeficiency virus type 1 variants (184V) require multiple amino acid changes to become co-resistant to zidovudine in vivo.

Exposure of human immunodeficiency virus to the nucleoside analogue lamivudine (3TC) rapidly selects for resistant variants with a valine at codon 184 (M184V) in the catalytic site of reverse transcriptase. In vitro, 184V demonstrated increased enzyme fidelity and suppressed zidovudine resistance. Clinical trials demonstrated that 3TC-zidovudine combination therapy results in a strong and sustained antiviral response. To investigate the role of 184V on in vivo virus evolution, the effect of zidovudine addition in 3TC-pretreated patients harboring 184V was studied. In vivo, no significant change in fidelity was observed with 184V, shown by generation of the classical pattern of zidovudine mutations. Of interest, in contrast to zidovudine monotherapy, in which just one substitution is sufficient for in vivo development of significant zidovudine resistance, multiple substitutions are required for the same level of zidovudine resistance in strains harboring 184V. This need for multiple substitutions may be one of the mechanisms explaining the sustained antiretroviral response of the 3TC-zidovudine combination.

Amino Acid Sequence↗

The origin and evolution of Ebola and Marburg viruses.

Molecular evolutionary analyses for Ebola and Marburg viruses were conducted with the aim of elucidating evolutionary features of these viruses. In particular, the rate of nonsynonymous substitutions for the glycoprotein gene of Ebola virus was estimated to be, on the average, 3.6 x 10(-5) per site per year. Marburg virus was also suggested to be evolving at a similar rate. Those rates were a hundred times slower than those of retroviruses and human influenza A virus, but were of the same order of magnitude as that of the hepatitis B virus. When these rates were applied to the degree of sequence divergence, the divergence time between Ebola and Marburg viruses was estimated to be more than several thousand years ago. Moreover, most of the nucleotide substitutions were transitions and synonymous for Marburg virus. This suggests that purifying selection has operated on Marburg virus during evolution.

DNA, Viral↗

Calicivirus infections in children.

PURPOSE OF REVIEW: Caliciviruses are a major cause of human illness, and are listed as category B pathogens according to the National Institute of Allergy and Infectious Diseases classification of pathogens important for biodefense. Caliciviruses are commonly encountered in contaminated food and water, and a large variety has been implicated as sources of infection during outbreak investigations. RECENT FINDINGS: New names for two of the four genera of the Caliciviridae were approved in 2002. They are Norovirus, for what were previously called Norwalk-like viruses or small, round-structured viruses, and Sapovirus, for what were previously called Sapporo-like viruses. Caliciviruses are highly diverse genetically and antigenically. This diversity complicates the design of diagnostic assays, yet can be used to discriminate contaminating and infecting strains during outbreak investigations. Of particular interest is the recent finding of naturally occurring recombinant Norovirus strains, all of which have been virulent and are widely dispersed and apparently ecologically indistinguishable from other calicivirus strains. This finding is considered in light of the evidence for recombination between caliciviruses and picornaviruses, and recombination as a more general phenomenon for virus evolution. SUMMARY: Continued investigations of calicivirus outbreaks are now focusing on the implicated sources of infection. While many foods and environmental waters have long been implicated as outbreak sources, the methods for detecting caliciviruses are being developed and refined. Recognition is now turning to unexpected sources of contamination, such as presumably clean foods and waters, including bottled water and minimally handled foods. Parallels between Norovirus and Salmonella ecology and epidemiology are noted, as a guide to understanding evolving new information about caliciviruses.

Caliciviridae↗

Rotavirus strain surveillance in Latin America: a review of the last nine years.

BACKGROUND: Latin America will likely be the first area in the developing world where rotavirus vaccine will be introduced into the routine childhood immunization schedule. In anticipation of that goal, we reviewed the distribution of group A rotavirus genotypes in Latin America to understand the diversity of strains to be targeted by vaccines and to identify novel strains that may pose challenges for vaccines. METHODS: We reviewed studies characterizing rotavirus strains in Latin America (published in English since 1995) that used molecular methods to type genes encoding the G and P outer capsid proteins, VP7 and VP4, and that reported data on >50 specimens. RESULTS: Fifteen studies from 5 countries met our criteria. In total, 1989 samples were characterized; 12% (233) were mixed rotavirus infections with more than 1 strain, and 20% (402) were not fully typable. Of the remaining 1354 samples that were fully typed, 83% represented the 4 common strains: P[8],G1 (40%); P[4],G2 (30%); P[8],G3 (6%); P[8],G4 (7%). The unusual strains provide interesting insights into virus evolution: some strains (G5) were regionally common; the emerging G9 strains were widely distributed; many animal-human reassortants were present; and some common serotypes (G3 and G4) were of animal origin. Also an unusual G12 serotype was recently detected in Argentina. CONCLUSIONS: The common rotavirus serotypes should remain the prime targets for vaccine development. However, the changing profile of rare strains, animal-human reassortants and nontypable strains suggest that rotavirus is constantly evolving. Laboratory surveillance is needed to monitor rotavirus strains now in circulation and to detect those that might escape the immunity induced by vaccines or represent vaccine strains entering the environment.

Capsid Proteins↗

The origin of acquired immune deficiency syndrome: Darwinian or Lamarckian?

The subtypes of human immunodeficiency virus type 1 (HIV-1) group M exhibit a remarkable similarity in their between-subtype distances, which we refer to as high synchrony. The shape of the phylogenetic tree of these subtypes is referred to as a sunburst to distinguish it from a simple star phylogeny. Neither a sunburst pattern nor a comparable degree of symmetry is seen in a natural process such as in feline immunodeficiency virus evolution. We therefore have undertaken forward-process simulation studies employing coalescent theory to investigate whether such highly synchronized subtypes could be readily produced by natural Darwinian evolution. The forward model includes both classical (macro) and molecular (micro) epidemiological components. HIV-1 group M subtype synchrony is quantified using the standard deviation of the between-subtype distances and the average of the within-subtype distances. Highly synchronized subtypes and a sunburst phylogeny are not observed in our simulated data, leading to the conclusion that a quasi-Lamarckian, punctuated event occurred. The natural transfer theory for the origin of human acquired immune deficiency syndrome (AIDS) cannot easily be reconciled with these findings and it is as if a recent non-Darwinian process took place coincident with the rise of AIDS in Africa.

Acquired Immunodeficiency Syndrome↗

Phylogenetic analysis of GB viruses A and C: evidence for cospeciation between virus isolates and their primate hosts.

GB viruses A and C (GBV-A and GBV-C) have been isolated from humans and non-human primates. Phylogenetic analysis based on full-length polyproteins suggests that these two viruses have a common ancestor. It has now been determined that analysis of subgenomic amino acid sequences in the E2 and NS5 regions of GBV-A and a 345 nucleotide segment in the 5' non-coding (5'NC) region was able to reproduce the phylogenetic relationships obtained by complete polyprotein sequences analysis. Using 5'NC sequences from databases, GBV-A isolates were discriminated into eight genetic groups, each one closely associated with specific primate hosts. Phylogenetic analyses performed on sequences from the epsilon-globin genes of primate hosts on one hand and complete polyprotein sequences from GBV-A and GBV-C isolates on the other suggest that a mechanism of cospeciation could be involved in virus evolution over a period of 35 million years.

Animals↗

In vitro analysis of human immunodeficiency virus type 1 resistance to nevirapine and fitness determination of resistant variants.

Nevirapine-resistant variants were generated by serial passages in MT-2 cells in the presence of increasing drug concentrations. In passage 5, mutations V106A, Y181C and G190A were detected in the global population, associated with a 100-fold susceptibility decrease. Sequence analysis of biological clones obtained from passage 5 and subsequent passages showed that single mutants, detected in first passages, were progressively replaced in passage 15 by double mutants, correlating with a 500-fold increase in phenotypic resistance. Fitness determination of single mutants confirmed that, in the presence of nevirapine, every variant was more fit than wild-type with a fitness order Y181C>V106A>G190A>wild-type. Unexpectedly, in the absence of the drug, the Y181C resistant mutant was more fit than wild-type, with a fitness gradient Y181C>wild-type >G106A>or=V190A. Using a molecular clone in which the Y181C mutation was introduced by in vitro mutagenesis, the greater fitness of the Y181C mutant was confirmed in new competition cultures. These data exemplify the role of resistance mutations on virus phenotype but also on virus evolution leading, occasionally, to resistant variants fitter than the wild-type in the absence of the drug.

Adaptation, Physiological↗

Viruses, plasmids and other genetic elements of thermophilic and hyperthermophilic Archaea.

We review and update the work on genetic elements, e.g., viruses and plasmids (exluding IS elements and transposons) in the kingdom Crenarchaeota (Thermoproteales and Sulfolobales) and the orders Thermococcales and Thermoplasmales in the kingdom Euryarchaeota of the archael domain, including unpublished data from our laboratory. The viruses of Crenarchaeota represent four novel virus families. The Fuselloviridae represented by SSVI of S. shibatae and relatives in other Sulfolobus strains have the form of a tailed spindle. The envelope is highly hydrophobic. The DNA is double-stranded and circular. Members of this group have also been found in Methanococcus and Haloarcula. The Lipothrivciridae (e.g., T TV1 to 3) have the form of flexible filaments. They have a core containing linear double-stranded DNA and DNA-binding proteins which is wrapped into a lipid membrane. The "Bacilloviridae" (e.g., TTV4 and SIRV) are stiff rods lacking this membrane, but also featuring linear double-stranded DNA and DNA-binding proteins. Both virus types carry on both ends structures involved in the attachment to receptors. Both types are represented in Thermoproteus and Sulfolobus. The droplet-formed novel Sulfolobus virus SNDV represents the "Guttaviridae" containing circular double-stranded DNA. Though head and tail viruses distantly resembling T phages or lambdoid phages were seen electronmicroscopically in solfataric water samples, no such virus has so far been isolated. SSV1 is temperate, TTV1 causes lysis after induction, the other viruses found so far exist in carrier states. The hosts of all but TTV1 survive virus production. We discuss the implications of the nature of these viruses for understanding virus evolution. The plasmids found so far range in size from 4.5 kb to about 40 kb. Most of them occur in high copy number, probably due to the way of their detection. Most are cryptic, pNOB8 is conjugative, the widespread pDL10 alleviates in an unknown way autotrophic growth of its host Desulfurolobus by sulfur reduction. The plasmid pTIK4 appears to encode a killer function. pNOB8 has been used as a vector for the transfer of the lac S (beta-galactosidase) gene into a mutant of S. solfataricus.

Archaea↗