Biomedical subjects

J Dopazo

Publications and source records attributed to J Dopazo.

At least 55 records · Page 3Linked to original sources

PubMed1994

Evolutionary relationships among bean common mosaic virus strains and closely related potyviruses.

Bean common mosaic virus (BCMV) consists of a large number of strains with complex and controversial relationships among them and with other potyviruses that infect legumes. In order to elucidate the BCMV taxonomic pattern and its evolutionary implications, a phylogenetic analysis has been carried out. The analysis of the coat protein gene and 3' non-coding region (NCR) sequences recently obtained by us and other currently available potyviral sequences confirms the clustering of viruses comprised in BCMV strains with other closely related potyviruses and reveals the great informative content of 3' NCR, suggesting a more relevant role for this region in phylogenetic analysis.

Amino Acid Sequence↗

PubMed1994

Evolutionary pattern of human respiratory syncytial virus (subgroup A): cocirculating lineages and correlation of genetic and antigenic changes in the G glycoprotein.

The genetic and antigenic variability of the G glycoproteins from 76 human respiratory syncytial (RS) viruses (subgroup A) isolated during six consecutive epidemics in either Montevideo, Uruguay, or Madrid, Spain, have been analyzed. Genetic diversity was evaluated for all viruses by the RNase A mismatch cleavage method and for selected strains by dideoxy sequencing. The sequences reported here were added to those published for six isolates from Birmingham, United Kingdom, and for two reference strains (A2 and Long), to derive a phylogenetic tree of subgroup A viruses that contained two main branches and several subbranches. During the same epidemic, viruses from different branches were isolated. In addition, closely related viruses were isolated in distant places and in different years. These results illustrate the capacity of the virus to spread worldwide, influencing its mode of evolution. The antigenic analysis of all isolates was carried out with a panel of anti-G monoclonal antibodies that recognized strain-specific (or variable) epitopes. A close correlation between genetic relatedness and antigenic relatedness in the G protein was observed. These results, together with an accumulation of amino acid changes in a major antigenic area of the G glycoprotein, suggest that immune selection may be a factor influencing the generation of RS virus diversity. The pattern of RS virus evolution is thus similar to that described for influenza type B viruses, expect that the level of genetic divergence among the G glycoproteins of RS virus isolates is the highest reported for an RNA virus gene product.

Amino Acid Sequence↗

PubMed1993

Split decomposition: a technique to analyze viral evolution.

A clustering technique allowing a restricted amount of overlapping and based on an abstract theory of coherent decompositions of finite metrics is used to analyze the evolution of foot-and-mouth disease viruses. The emerging picture is compatible with the existence of viral populations with a quasispecies structure and illustrates various forms of evolution of this virus family. In addition, it allows the correlation of these forms with geographic occurrence.

Animals↗

PubMed1993

Evolution and tropism of transmissible gastroenteritis coronavirus.

Transmissible gastroenteritis coronavirus (TGEV) is an enteropathogenic coronavirus isolated for the first time in 1946. Nonenteropathogenic porcine respiratory coronaviruses (PRCVs) have been derived from TGEV. The genetic relationship among six European PRCVs and five coronaviruses of the TGEV antigenic cluster has been determined based on their RNA sequences. The S proteins of six European PRCVs have an identical deletion of 224 amino acids starting at position 21. The deleted area includes the antigenic sites C and B of TGEV S glycoprotein. Interestingly, two viruses (NEB72 and TOY56) with respiratory tropism have the S protein with a similar size to the enteric viruses. NEB72 and TOY56 viruses have 2 and 15 specific amino acid differences with the enteric viruses, respectively. Four of the residues changed are located within the deletion present in the PRCVs and may influence the enteric tropism of TGEV in vivo. A receptor binding site (RBS) used by the virus to infect ST and other cell types might be located between sites A and D of the S glycoprotein, since monoclonal antibodies (MAbs) specific for these sites inhibit the binding of the virus to ST cells. An evolutionary tree relating 13 enteric and respiratory isolates has been proposed. According to this tree, a main virus lineage evolved from a recent progenitor which was circulating around 1941. From this, secondary lineages originated PUR46, NEB72, TOY56, MIL65, BRI70, and the PRCVs, in this order. Least squares estimation of the origin of TGEV-related coronaviruses showed a significant constancy in the mutation fixation rate.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

PubMed1993

A computer program for the design of PCR primers for diagnosis of highly variable genomes.

PCRDiag (Diagnosis by PCR) is a computer program which allows the localization of pairs of oligonucleotides with optimal thermodynamic requirements for use in a PCR assay. The program is designed for the selection of pairs of primers complementary to sequences present in a group, whose identification is intended, but are absent in other non-specific sequences. The program constitutes a powerful tool, specially in systems which display a high degree of sequence heterogeneity, as is the case of RNA viruses. The program runs on IBM-PC and compatible computers and has no special software requirements. It does not need the previous alignment of the sequences analyzed.

Algorithms↗

PubMed1993

Estimates by computer simulation of genetic distances from comparisons of RNAse A mismatch cleavage patterns.

The RNAse A mismatch cleavage method was used to analyze genomic variability in RNA and DNA systems. However, there is no method which relates the digestion patterns observed to the extent of genetic variation. Here we report computer simulations which provide a simple estimator of genetic distances from the comparison of RNAse A digestion patterns. The results show that the number of non-shared fragments is proportional to the number of mutations between each pair of sequences compared. This prediction is supported by the comparison of the RNAse A mismatch patterns and the nucleotide sequences of a set of influenza A (H3N2) hemagglutinin genes. The procedure allows a quantitative and reliable use of the RNAse A mismatch cleavage method.

Computer Simulation↗

PubMed1993

Analysis of genetic variability of populations of herpes simplex viruses.

An analysis of genetic variability of herpes simplex virus type 1 and type 2 populations of the Madrid (Spain) area has been carried out by digestion of viral DNA with restriction endonucleases. The index of nucleotide diversity indicated that herpes simplex virus type 1 has a slightly, although statistically significant, higher degree of heterogeneity than type 2. A phylogenetic tree for each type of virus has been constructed. The evolutionary pattern followed by both types of viruses ('star-like' topology) suggest that all the isolates analyzed evolved from a unique origin for each type of virus.

Animals↗

PubMed1993

Quantitative characterization of antigens using monoclonal antibody reactivities.

A multipurpose program that empirically relates antigenic reactivities with monoclonal antibodies (MAbs) to genetic distances is presented. The program uses a set of known genetic pairwise distances to weigh each MAb depending on its capacity to define groups of taxonomically related antigens. This allows highly accurate identification and classification of unknown antigens. Also, the weights obtained constitute a quantitative measure of epitope conservation and can be used for improved vaccine design.

Algorithms↗

PubMed1993

Design of primers for PCR amplification of highly variable genomes.

A program to aid in the search of primers for specific polymerase chain reaction (PCR) amplification of highly variable genomes is presented. It involves the derivation of variability profiles to identify optimal regions for PCR amplification, taking into account stability of DNA-primer hybrids. An application of the program to foot-and-mouth disease virus diagnosis is presented.

Aphthovirus↗

PubMed1993

Estimation of the intrinsic rate of natural increase and its error by both algebraic and resampling approaches.

The intrinsic rate of natural increase or Malthusian parameter plays a key role fields as diverse as ecology, genetics, demography and evolution. It characterizes the growth of a population in a determinate environment. Since its rigorous statistical estimation requires of intensive calculation, the use of a computer becomes essential. The two main approaches to the calculation of the Malthusian parameter, its error and confidence intervals have been implemented in a program and have been compared by means of an example.

Algorithms↗

PubMed1993

Molecular epidemiology of foot-and-mouth disease virus type O.

A phylogenetic tree based on the VPI sequences of type O foot-and-mouth disease virus (FMDV) has been derived. Direct sequencing of PCR products has been used to obtain the VP1 gene sequences of new isolates. The tree exhibits four main lineages that largely correlate with the geographical origin of isolates. The analysis supports a close relationship between European O1 field isolates and vaccine strains, with the exception of O Thalheim Aus/81 and O Wuppertal Ger/82 which were probably of non-European origin. Analysis of nucleotide substitutions indicates that synonymous mutations play a major role in FMDV evolution.

Amino Acid Sequence↗

PubMed1992

Phylogenetic analysis of the thiolase family. Implications for the evolutionary origin of peroxisomes.

The thiolase family is a widespread group of proteins present in prokaryotes and three cellular compartments of eukaryotes. This fact makes this family interesting in order to study the evolutionary process of eukaryotes. Using the sequence of peroxisomal thiolase from Saccharomyces cerevisiae recently obtained by us and the other known thiolase sequences, a phylogenetic analysis has been carried out. It shows that all these proteins derived from a primitive enzyme, present in the common ancestor of eubacteria and eukaryotes, which evolved into different specialized thiolases confined to various cell compartments. The evolutionary tree obtained is compatible with the endosymbiotic theory for the origin of peroxisomes.

Acetyl-CoA C-Acetyltransferase↗

PubMed1992

Primer design for specific diagnosis by PCR of highly variable RNA viruses: typing of foot-and-mouth disease virus.

A PCR assay for the specific detection and identification of viral sequences that correlate with established serotypes of foot-and-mouth disease virus (FMDV) has been developed. A new analysis based on homology profiles among reported sequences was used for primer design. RNA replicase (3D) gene regions that showed high homology among FMDVs, and low homology to other picornaviruses, were used for PCR amplification. Specific and highly sensitive detection was achieved for RNA of FMDV types C, A, and O, either purified or extracted from vesicular fluids of infected animals, under reaction conditions permissive for the detection of variants present in the virus population. Similarly, serotype-specific primers were designed to amplify the carboxy-terminal end of VP1 gene of FMDV types either C, A, or O. The results of PCR amplification of 15 different FMDV RNAs using type-specific primers are in agreement with the serological typing of the corresponding viruses and show that the primer-selection procedure developed for FMDV constitutes a reliable method of viral diagnosis.

Animals↗

PubMed1992

Genetic evolution and tropism of transmissible gastroenteritis coronaviruses.

Transmissible gastroenteritis virus (TGEV) is an enteropathogenic coronavirus isolated for the first time in 1946. Nonenteropathogenic porcine respiratory coronaviruses (PRCVs) have been derived from TGEV. The genetic relationship among six European PRCVs and five coronaviruses of the TGEV antigenic cluster has been determined based on their RNA sequences. The S protein of six PRCVs have an identical deletion of 224 amino acids starting at position 21. The deleted area includes the antigenic sites C and B of TGEV S glycoprotein. Interestingly, two viruses (NEB72 and TOY56) with respiratory tropism have S proteins with a size similar to the enteric viruses. NEB72 and TOY56 viruses have in the S protein 2 and 15 specific amino acid differences with the enteric viruses. Four of the residues changed (aa 219 of NEB72 isolate and aa 92, 94, and 218 of TOY56) are located within the deletion present in the PRCVs and may be involved in the receptor binding site (RBS) conferring enteric tropism to TGEVs. A second RBS used by the virus to infect ST cells might be located in a conserved area between sites A and D of the S glycoprotein, since monoclonal antibodies specific for these sites inhibit the binding of the virus to ST cells. An evolutionary tree relating 13 enteric and respiratory isolates has been proposed. According to this tree, a main virus lineage evolved from a recent progenitor virus which was circulating around 1941. From this, secondary lineages originated PUR46, NEB72, TOY56, MIL65, BR170, and the PRCVs, in this order. Least squares estimation of the origin of TGEV-related coronaviruses showed a significant constancy in the fixation of mutations with time, that is, the existence of a well-defined molecular clock. A mutation fixation rate of 7 +/- 2 x 10(-4) nucleotide substitutions per site and per year was calculated for TGEV-related viruses. This rate falls in the range reported for other RNA viruses. Point mutations and probably recombination events have occurred during TGEV evolution.

Amino Acid Sequence↗

PubMed1992

Evolution of the capsid protein genes of foot-and-mouth disease virus: antigenic variation without accumulation of amino acid substitutions over six decades.

The genetic diversification of foot-and-mouth disease virus (FMDV) of serotype C over a 6-decade period was studied by comparing nucleotide sequences of the capsid protein-coding regions of viruses isolated in Europe, South America, and The Philippines. Phylogenetic trees were derived for VP1 and P1 (VP1, VP2, VP3, and VP4) RNAs by using the least-squares method. Confidence intervals of the derived phylogeny (significance levels of nodes and standard deviations of branch lengths) were placed by application of the bootstrap resampling method. These procedures defined six highly significant major evolutionary lineages and a complex network of sublines for the isolates from South America. In contrast, European isolates are considerably more homogeneous, probably because of the vaccine origin of several of them. The phylogenetic analysis suggests that FMDV CGC Ger/26 (one of the earliest FMDV isolates available) belonged to an evolutionary line which is now apparently extinct. Attempts to date the origin (ancestor) of the FMDVs analyzed met with considerable uncertainty, mainly owing to the stasis noted in European viruses. Remarkably, the evolution of the capsid genes of FMDV was essentially associated with linear accumulation of silent mutations but continuous accumulation of amino acid substitutions was not observed. Thus, the antigenic variation attained by FMDV type C over 6 decades was due to fluctuations among limited combinations of amino acid residues without net accumulation of amino acid replacements over time.

Amino Acid Sequence↗

PubMed1991

Fixation of mutations at the VP1 gene of foot-and-mouth disease virus. Can quasispecies define a transient molecular clock?

The number of nucleotide (nt) substitutions found in the VP1 gene (encoding viral capsid protein) between any two of 16 closely related isolates of foot-and-mouth disease virus (FMDV) has been quantified as a function of the time interval between isolations [Villaverde et al., J. Mol. Biol. 204 (1988) 771-776]. One of them (isolate C-S12) includes some replacements found in isolates that preceded it and other replacements found in later isolates. The study has revealed alternating periods of rapid evolution and of relative genetic stability of VP1. During a defined period of acute disease, the rate of fixation of replacements at the VP1 coding segment was 6 x 10(-3) substitutions per nt per year. Only small differences in the rate of evolution were observed between subsegments within the VP1 gene. The observation of a relatively constant rate of evolution during a disease episode was unexpected. We propose that such constancy may be a consequence of random sampling of mutants from the FMDV quasispecies, followed by their amplification in susceptible hosts (to generate a new quasispecies). Successive sampling and amplification events may result in a steady accumulation of mutations.

Amino Acid Sequence↗

PubMed1991

Phylogeny of viroids, viroidlike satellite RNAs, and the viroidlike domain of hepatitis delta virus RNA.

We report a phylogenetic study of viroids, some plant satellite RNAs, and the viroidlike domain of human hepatitis delta virus RNA. Our results support a monophyletic origin of these RNAs and are consistent with the hypothesis that they may be "living fossils" of a precellular RNA world. Moreover, the viroidlike domain of human hepatitis delta virus RNA appears closely related to the viroidlike satellite RNAs of plants, with which it shares some structural and functional properties. On the basis of our phylogenetic analysis, we propose a taxonomic classification of these RNAs.

Hepatitis Delta Virus↗

PubMed1990

Sequences of isopenicillin N synthetase genes suggest horizontal gene transfer from prokaryotes to eukaryotes.

Evolutionary distances between bacterial and fungal isopenicillin N synthetase (IPNS) genes have been compared to distances between the corresponding 5S rRNA genes. The presence of sequences homologous to the IPNS gene has been examined in DNAs from representative prokaryotic organisms and Ascomycotina. The results of both analyses strongly support two different events of horizontal transfer of the IPNS gene from bacteria to filamentous fungi. This is the first example of such a type of transfer from prokaryotes to eukaryotes.

Acremonium↗