Genetic and Phylogenetic Analysis of Glycoprotein of Rabies Virus Isolated from Several Species in Brazil

Sato, Go; Itou, Takuya; Shoji, Youko; Miura, Yasuo; Mikami, Takeshi; Ito, Makoto; Kurane, Ichiro; Samara, Samir Issa; Carvalho, Adolorata Aparecida Bianco; Nociti, Darci P.; Ito, Fumio; Sakai, Takeo

doi:10.1292/jvms.66.747

Cited by 54 publications

(41 citation statements)

References 20 publications

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“…Phylogenetic analyses based on the P and M genes revealed the existence of several Brazilian RV variants corresponding to phylogenetic analyses targeting the viral N and G genes [19,25,26,41,42,44,45]. Phylogenetic analysis showed two clusters consisting of chiroptera-or carnivora-related RV isolates.…”

Section: Discussionmentioning

confidence: 99%

“…RT-PCR and sequencing: RT-PCR and sequencing were performed as previously described [41]. The primers used for RT-PCR and sequencing for complete P and M genes are shown in Table 2.…”

Section: Virusesmentioning

confidence: 99%

“…sp.) have frequently been typed as vampire bat-related RVs in Brazil [19,25,26,41,42,44,45]. Variability has also been reported in the G protein, a major contributor to the pathogenicity of the virus, in which several amino acid substitutions at antigenic sites associated with the pathogenicity and immunogenicity of the virus have been identified in Brazilian RV variants [41].…”

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confidence: 99%

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Genetic Analysis of Phosphoprotein and Matrix Protein of Rabies Viruses Isolated in Brazil

Kobayashi

Okuda

Nakamura

et al. 2007

The Journal of Veterinary Medical Science

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ABSTRACT. To investigate the genetic characteristics of phosphoprotein (P) and matrix protein (M) genes of variable rabies virus (RV) prevalent in Brazil, the authors genetically characterized the P and M genes from 30 Brazilian RV field isolates. Phylogenetic analysis based on the P and M genes revealed the presence of six RV variants that consisted primarily of three insectivorous bats, the vampire bat, dog and fox in Brazil. Specific amino acid substitutions corresponding to these phylogenetic lineages were observed, with Asp 42 and Glu 62 in the P protein found to be characteristic of Brazilian chiroptera-and carnivora-related RVs, respectively. Amino acid sequence motifs predicted to associate with a viral function in the P and M proteins were conserved among Brazilian RV variants. KEY WORDS: Brazil, genetic analysis, matrix protein, phosphoprotein, rabies virus.J. Vet. Med. Sci. 69(11): 1145-1154 Rabies virus (RV) is a member of the Lyssavirus genus, which belongs to the Rhabdoviridae family. Lyssavirus is characterized as having seven genotypes (GTs) comprising, rabies virus (GT 1), Lagos bat virus (GT 2), Mokola virus (GT 3), Duvenhage virus (GT 4), European bat lyssavirus type (EBL) 1 (GT 5), EBL 2 (GT 6) and Australian bat lyssavirus (GT 7). Lyssaviruses have approximately 12-kb of unsegmented negative-stranded genomic RNA for encoding the genes of the nucleoprotein (N), phosphoprotein (P), matrix protein (M), glycoprotein (G) and polymerase (L).RV has an almost global distribution and infects a wide range of mammalian species in which it causes a lethal form of encephalopathy. In Brazil, the principal RV transmitters to humans and domestic animals -dogs and vampire batshave caused serious problems in the public health sector and the livestock industry [6,19]. In addition, RVs have been isolated from other animal species [8]. Previously, phylogenetic analyses targeting the N and G genes revealed that there were several RV variants which varied depending on the host species, which included vampire bats, insectivorous bats, dogs and foxes. Furthermore, RV isolates from cattle and frugivorous bats (Artibeus (A.) sp.) have frequently been typed as vampire bat-related RVs in Brazil [19,25,26,41,42,44,45]. Variability has also been reported in the G protein, a major contributor to the pathogenicity of the virus, in which several amino acid substitutions at antigenic sites associated with the pathogenicity and immunogenicity of the virus have been identified in Brazilian RV variants [41]. However, differences in the pathogenicity and antigenic characteristics of these Brazilian RV variants are not yet known. Recently, in addition to the G protein, both P and M proteins have also been reported to be associated with the pathogenicity of the virus [43]. The P protein forms a ribonucleoprotein (RNP) with N and L proteins, which then wraps around the viral RNA, and plays an important role in transcription and replication in conjunction with the L protein [3,5,11]. In addition, the P protein acts to counteract the ...

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Section: Discussionmentioning

confidence: 99%

“…RT-PCR and sequencing: RT-PCR and sequencing were performed as previously described [41]. The primers used for RT-PCR and sequencing for complete P and M genes are shown in Table 2.…”

Section: Virusesmentioning

confidence: 99%

mentioning

confidence: 99%

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Genetic Analysis of Phosphoprotein and Matrix Protein of Rabies Viruses Isolated in Brazil

Kobayashi

Okuda

Nakamura

et al. 2007

The Journal of Veterinary Medical Science

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“…The RT-PCR and RT-PCR products purification were performed according to our previously study [16]. P1/N8 and P1/BRABP-C1 primer pairs specifically amplified 1,482 and 1,430 nucleotide sequences of RV N gene, respectively.…”

Section: Rt-pcrmentioning

confidence: 99%

“…Direct sequencing and phylogenetic analysis: Direct sequencing, connection of obtained nucleotide fragments, design of a phylogenetic tree and calculation of nucleotide homologies were performed as follows our previous study [16]. Nucleotide sequences and the position of primers used for direct sequencing were shown in Table 2.…”

Section: Al4 Br-nl1 Br-mm1 Br-ma1 Br-ef2 Br-ef3 Br-bat1 and Br-mentioning

confidence: 99%

Molecular Epidemiological Analysis of Bat Rabies Viruses in Brazil

Kobayashi

Sato

Shoji

et al. 2005

The Journal of Veterinary Medical Science

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ABSTRACT. A molecular epidemiological analysis was performed in 19 rabies viruses (RVs) isolated from haematophagous, frugivorous and insectivorous bats, in Sao Paulo, Brazil. The authors carried out RT-PCR for amplification of the RV nucleoprotein (N) gene, and determined 1,335 nucleotide sequences of N gene by direct sequencing method. Phylogenetic analysis, which was based on the N gene of Brazilian RV isolates identified presently and previously, revealed that RVs isolated from bats were genetically divided into four lineages had a tendency to depend on the host bat species. Rabies viruses (RVs) isolated in Brazil belong to genotype 1 of genus lyssavirus, family Rhabdoviridae. Ito et al. [7,8] reported that main reservoirs of RVs were dogs and vampire bats and that the RVs were classified into dogrelated RVs and vampire bat-related RVs in Brazil. In some areas of Brazil, the rabies outbreaks by both dog-and batrelated RVs occurred at the same time [7]. RVs have been isolated from not only haematophagous bats but also frugivorous and insectivorous bats in Brazil. In Argentina, RVs isolated from frugivorous bats (Artibeus lituratus) were identified to be the variant of RV associated with haematophagous bats by use of the monoclonal antibody panels [4]. Recently we have demonstrated that RVs isolated from Brazilian frugivorous bats (Artibeus spp.) were phylogenetically characterized as the vampire bat-related RV [18].There are many reports of human cases on infection with insectivorous bat RVs, and the number of the cases tend to increase in North America [10][11][12]. In 2002, Favi et al. [5] reported that insectivorous bat (Tadarida brasiliensis) RV variant was isolated from a human patient in Chile. In Colombia and Venezuela insectivorous bat RV variants were also isolated from dogs and cats, which were main RVtransmitters to human [2,14]. Although some kinds of insectivorous bats have been diagnosed as RV-positive in Brazil, there have been no reports of the human case on infection with insectivorous bat RV variants until now. RVpositive insectivorous bats were, however, found in urban areas, and this fact has been considered as important problems in public health [1,6,17,21].As there is little information about the genetic characteristics of bat-related RVs in Brazil, authors determined nucleotide sequences on the N gene of RVs isolated from 9 Brazilian bat species. Phylogenetic analysis was also performed for the epidemiological study with the isolates identified previously. MATERIALS AND METHODS Viruses:In this study, 19 RVs were isolated from 3 haematophagous bats (Desmodus rotundus), 1 frugivorous bat (Artibeus lituratus), 13 insectivorous bats (3 Nyctinomops laticaudatus, 2 Eumops auripendulus, 4 Eptesicus furinalis, 2 Molossus molossus, 1 Molossus abrasus and 1 Molossus rufus) and 2 unclassified bats, and their nucleoprotein gene sequences were determined. Brain materials from these bats were diagnosed as RV-positive by immunofluorescent antibody test (FAT) using a rabbit polyclonal anti-rabies nucle...

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Functional identification of glutamate cysteine ligase and glutathione synthetase in the marine yeast Rhodosporidium diobovatum

Kong

Wang

Tian

et al. 2017

Sci Nat

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Glutathione (GSH) fulfills a variety of metabolic functions, participates in oxidative stress response, and defends against toxic actions of heavy metals and xenobiotics. In this study, GSH was detected in Rhodosporidium diobovatum by high-performance liquid chromatography (HPLC). Then, two novel enzymes from R. diobovatum were characterized that convert glutamate, cysteine, and glycine into GSH. Based on reverse transcription PCR, we obtained the glutathione synthetase gene (GSH2), 1866 bp, coding for a 56.6-kDa protein, and the glutamate cysteine ligase gene (GSH1), 2469 bp, coding for a 90.5-kDa protein. The role of GSH1 and GSH2 for the biosynthesis of GSH in the marine yeast R. diobovatum was determined by deletions using the CRISPR-Cas9 nuclease system and enzymatic activity. These results also showed that GSH1 and GSH2 were involved in the production of GSH and are thus being potentially useful to engineer GSH pathways. Alternatively, pET-GSH constructed using vitro recombination could be used to detect the function of genes related to GSH biosynthesis. Finally, the fermentation parameters determined in the present study provide a reference for industrial GSH production in R. diobovatum.

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Genetic and Phylogenetic Analysis of Glycoprotein of Rabies Virus Isolated from Several Species in Brazil

Cited by 54 publications

References 20 publications

Genetic Analysis of Phosphoprotein and Matrix Protein of Rabies Viruses Isolated in Brazil

Genetic Analysis of Phosphoprotein and Matrix Protein of Rabies Viruses Isolated in Brazil

Molecular Epidemiological Analysis of Bat Rabies Viruses in Brazil

Functional identification of glutamate cysteine ligase and glutathione synthetase in the marine yeast Rhodosporidium diobovatum

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