Sequencing and In Silico Multi-aspect Analysis of S1 Glycoprotein in 793/B Serotype of Infectious Bronchitis Virus Isolated From Iran in 2003 and 2011

Document Type: Original Articles

Authors

1 Department of Poultry Diseases, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran

2 Department of Animal Virology, Research and Diagnosis, Razi Vaccine and Serum Research Institute, Agricultural Research, Education, and Extension Organization, Karaj, Iran

3 Department of Research, Breeding and Production of Laboratory Animals, Razi Vaccine and Serum Research Institute, Agricultural Research, Education, and Extension Organization, Karaj, Iran

4 Department of Brucellosis, Razi Vaccine and Serum Research Institute, Agricultural Research, Education, and Extension Organization, Karaj, Iran

Abstract

Infectious bronchitis (IB) is an acute, highly contagious, and economically important viral disease of chickens. The S1 subunit from Spike (S) protein plays the major role in protective immunity and is involved in the host-virus interactions, as well as infectious bronchitis virus (IBV) serotyping. Aim of the present study was multi-aspect analysis of the molecular and immunological features of 5' part belonging to the S1 glycoprotein sequence of Iranian 793/B IBV strain isolates. This might ideally help in characterization, prevention, and vaccine development. The tissue samples were prepared, followed by virus isolation, reverse transcription polymerase chain reaction and restriction fragment length polymorphism analysis. In addition, sequencing and registration of the sequences in the National Center for Biotechnology Information were performed. Moreover, 12 sequences were retrieved from Fars province, Iran. The next steps included evaluation of conservation/variability along the sequences, phylogenetic analysis, estimation of the average evolutionary divergence over all the sequence pairs, predicting the phosphorylation/N-glycosylation/palmitoylation sites, and the final analysis of antigenicity. The findings of alignment, entropy plot, and pairwise similarity analysis revealed 17 hypervariable regions. The isolates belonging to Tehran were clustered in phylogenetic tree, and the most similar isolates to them were ADW11182 and ADW11183. Location of some of the N-glycosylation/phosphorylation/palmitoylation points indicated that these sites were conserved among the isolates. Furthermore, the frequency of epitopes and their scores reflect the high immunogenicity of S1 protein in 793/B serotype. Analysis of the primary and secondary structures demonstrated that their parameters had variable values and were different regarding the number and location of α-helix, β-strand, and coils. According to our findings, the Iranian isolates of 793/B serotype change their molecular characteristics during time and in different geographical regions. These alterations might account for failure in prevention programs and differences in virulence and pathogenicity.

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Main Subjects


Article Title [French]

Séquençage et Analyse in Silico d’Aspects Multiples de la Glycoprotéine S1 dans le Sérotype 793 / B du de la Bronchite Infectieuse Isolé en l'Iran dans les Années 2003 et 2011

Abstract [French]

La bronchite infectieuse (BI) est une maladie virale aiguë, hautement contagieuse et économiquement importante chez les poulets. La sous-unité S1 de la protéine Spike (S) joue un rôle majeur dans l'immunité protectrice et est impliquée dans les interactions hôte-virus, ainsi que dans le sérotypage du virus de la bronchite infectieuse (IBV). Le but de cette étude était l'analyse de plusieurs aspects des caractéristiques moléculaires et immunologiques de la partie 5 'appartenant à la séquence glycoprotéique S1 d'isolats de souche IBV 793 / B iraniens. Cela pourrait idéalement aider à la caractérisation, à la prévention et au développement de vaccins. Les échantillons de tissu ont été préparés, le virus a été ensuite isol, et une réaction en chaîne par polymérase en transcription inverse a a été menée pour l'analyse du polymorphisme de la longueur des frragments de restriction. De plus, le séquençage et l'enregistrement des séquences ont été effectués dans le Centre national d'information sur la biotechnologie. séquencesDouze séquences ont été récupérées de la province de Fars, en Iran. Les étapes suivantes comprenaient l'évaluation de la conservation / variabilité des séquences, l'analyse phylogénétique, l'estimation de la divergence évolutive moyenne sur toutes les paires de séquences, la prédiction des sites de phosphorylation / N-glycosylation / palmitoylation et l'analyse finale de l'antigénicité. Les résultats de l'alignement, du tracé d'entropie et de l'analyse de similarité par paires ont révélé 17 régions hypervariables. Les isolats àprovenant de Téhéran regroupés dans un arbre phylogénétique et les isolats les plus similaires étaient ADW11182 et ADW11183. L'emplacement de certains des points de N-glycosylation / phosphorylation / palmitoylation indique que ces sites sont conservés parmi les isolats. De plus, la fréquence des épitopes et leurs scores reflètent l'immunogénicité élevée de la protéine S1 dans le sérotype 793 / B. L'analyse des structures primaires et secondaires a montré que leurs paramètres avaient des valeurs variables et étaient différents pour le nombre et la localisation de l'hélice α, du brin β et des spires. Selon nos résultats, les isolats iraniens de sérotype 793/ B modifient leurs caractéristiques moléculaires au cours du temps et selon les différentes régions géographiques. Ces modifications pourraient expliquer l'échec des programmes de prévention et les différences de virulence et de pathogénicité.

Keywords [French]

  • Sérotype 793 / B
  • Bioinformatique
  • Virus de la bronchite infectieuse
  • Caractéristiques Moléculaires
  • Glycoprotéine S1
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