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Complete genomic sequence analysis of infectious bronchitis virus Ark DPI strain and its evolution by recombination.

Ammayappan A, Upadhyay C, Gelb J, Vakharia VN - Virol. J. (2008)

Bottom Line: The genome of Ark DPI consists of 27,620 nucleotides, excluding poly (A) tail, and comprises ten open reading frames.Possible recombination sites were found at the intergenic region of spike gene, 3'end of S1 and 3a gene.Independent recombination events may have occurred in the entire genome of Ark DPI, involving four different IBV strains, suggesting that genomic RNA recombination may occur in any part of the genome at number of sites.

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Affiliation: Center of Marine Biotechnology, University of Maryland Biotechnology Institute, Baltimore, 701 East Pratt Street, Baltimore, Maryland 21202-3101, USA. ammayapp@umbi.umd.edu

ABSTRACT
An infectious bronchitis virus Arkansas DPI (Ark DPI) virulent strain was sequenced, analyzed and compared with many different IBV strains and coronaviruses. The genome of Ark DPI consists of 27,620 nucleotides, excluding poly (A) tail, and comprises ten open reading frames. Comparative sequence analysis of Ark DPI with other IBV strains shows striking similarity to the Conn, Gray, JMK, and Ark 99, which were circulating during that time period. Furthermore, comparison of the Ark genome with other coronaviruses demonstrates a close relationship to turkey coronavirus. Among non-structural genes, the 5'untranslated region (UTR), 3C-like proteinase (3CLpro) and the polymerase (RdRp) sequences are 100% identical to the Gray strain. Among structural genes, S1 has 97% identity with Ark 99; S2 has 100% identity with JMK and 96% to Conn; 3b 99%, and 3C to N is 100% identical to Conn strain. Possible recombination sites were found at the intergenic region of spike gene, 3'end of S1 and 3a gene. Independent recombination events may have occurred in the entire genome of Ark DPI, involving four different IBV strains, suggesting that genomic RNA recombination may occur in any part of the genome at number of sites. Hence, we speculate that the Ark DPI strain originated from the Conn strain, but diverged and evolved independently by point mutations and recombination between field strains.

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Classical Genome Organization of IBV-Ark DPI. The genome of Ark DPI is 27,620 nt long, excluding poly (A) tract. Middle: ten genes and its ORFs. Ribosomal frameshift and position of transcriptional regulatory sequences (TRS) of each gene is indicated. Top: putative domains of ORF1a/1b polyprotein: nsp-non-structural protein, Ac-acidic domain, X-unknown domain X, PL1- papain like proteinase1, PL2-papain like proteinase 2; Y-unknown domain Y; HD-hydrophobic domain, 3CL-3C-like proteinase, G-Growth factor like protein, RdRp-RNA dependent RNA polymerase, Hel-helicase, ExoN-exoribonuclease, Ne-nidoviral uridylate-specific endoribonuclease, MT- 2'-O-ribose methyltransferase. Bottom: details of spike protein; SP-signal peptide, RRSRR/S- spike protein cleavage site between 544 and 545aa, TM-transmembrane domain of spike protein.
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Figure 1: Classical Genome Organization of IBV-Ark DPI. The genome of Ark DPI is 27,620 nt long, excluding poly (A) tract. Middle: ten genes and its ORFs. Ribosomal frameshift and position of transcriptional regulatory sequences (TRS) of each gene is indicated. Top: putative domains of ORF1a/1b polyprotein: nsp-non-structural protein, Ac-acidic domain, X-unknown domain X, PL1- papain like proteinase1, PL2-papain like proteinase 2; Y-unknown domain Y; HD-hydrophobic domain, 3CL-3C-like proteinase, G-Growth factor like protein, RdRp-RNA dependent RNA polymerase, Hel-helicase, ExoN-exoribonuclease, Ne-nidoviral uridylate-specific endoribonuclease, MT- 2'-O-ribose methyltransferase. Bottom: details of spike protein; SP-signal peptide, RRSRR/S- spike protein cleavage site between 544 and 545aa, TM-transmembrane domain of spike protein.

Mentions: The details of genome organization of Ark DPI are shown in Fig. 1. IBV polyprotein is cleaved into 15 cleavage products, among which first two N-terminal products are cleaved by PLpro and rest of the C-terminal products are cleaved by 3CLpro [15]. The putative domains and their cleavage sites (Fig. 1) are predicted by comparison of amino acid sequences of each non-structural protein (nsp) of Ark DPI with those of IBV-Beaudette which is available in Coronavirus Database (CoVDB) [16]. The nucleotide and the amino acid identity of Ark DPI with other IBVs and coronaviruses are listed in Tables 1, 2, 3. The whole structural gene of Jilin is 100% identical to Ark DPI, which suggests that Jilin strain is actually Ark DPI, which is currently used as a vaccine in China [17]. The whole genome comparison of IBV strains reveals a close relationship of Ark DPI with Cal 99 (96% identity), as shown in Fig. 2. Earlier studies have shown that Cal 99 probably evolved from Ark DPI [18].


Complete genomic sequence analysis of infectious bronchitis virus Ark DPI strain and its evolution by recombination.

Ammayappan A, Upadhyay C, Gelb J, Vakharia VN - Virol. J. (2008)

Classical Genome Organization of IBV-Ark DPI. The genome of Ark DPI is 27,620 nt long, excluding poly (A) tract. Middle: ten genes and its ORFs. Ribosomal frameshift and position of transcriptional regulatory sequences (TRS) of each gene is indicated. Top: putative domains of ORF1a/1b polyprotein: nsp-non-structural protein, Ac-acidic domain, X-unknown domain X, PL1- papain like proteinase1, PL2-papain like proteinase 2; Y-unknown domain Y; HD-hydrophobic domain, 3CL-3C-like proteinase, G-Growth factor like protein, RdRp-RNA dependent RNA polymerase, Hel-helicase, ExoN-exoribonuclease, Ne-nidoviral uridylate-specific endoribonuclease, MT- 2'-O-ribose methyltransferase. Bottom: details of spike protein; SP-signal peptide, RRSRR/S- spike protein cleavage site between 544 and 545aa, TM-transmembrane domain of spike protein.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2628353&req=5

Figure 1: Classical Genome Organization of IBV-Ark DPI. The genome of Ark DPI is 27,620 nt long, excluding poly (A) tract. Middle: ten genes and its ORFs. Ribosomal frameshift and position of transcriptional regulatory sequences (TRS) of each gene is indicated. Top: putative domains of ORF1a/1b polyprotein: nsp-non-structural protein, Ac-acidic domain, X-unknown domain X, PL1- papain like proteinase1, PL2-papain like proteinase 2; Y-unknown domain Y; HD-hydrophobic domain, 3CL-3C-like proteinase, G-Growth factor like protein, RdRp-RNA dependent RNA polymerase, Hel-helicase, ExoN-exoribonuclease, Ne-nidoviral uridylate-specific endoribonuclease, MT- 2'-O-ribose methyltransferase. Bottom: details of spike protein; SP-signal peptide, RRSRR/S- spike protein cleavage site between 544 and 545aa, TM-transmembrane domain of spike protein.
Mentions: The details of genome organization of Ark DPI are shown in Fig. 1. IBV polyprotein is cleaved into 15 cleavage products, among which first two N-terminal products are cleaved by PLpro and rest of the C-terminal products are cleaved by 3CLpro [15]. The putative domains and their cleavage sites (Fig. 1) are predicted by comparison of amino acid sequences of each non-structural protein (nsp) of Ark DPI with those of IBV-Beaudette which is available in Coronavirus Database (CoVDB) [16]. The nucleotide and the amino acid identity of Ark DPI with other IBVs and coronaviruses are listed in Tables 1, 2, 3. The whole structural gene of Jilin is 100% identical to Ark DPI, which suggests that Jilin strain is actually Ark DPI, which is currently used as a vaccine in China [17]. The whole genome comparison of IBV strains reveals a close relationship of Ark DPI with Cal 99 (96% identity), as shown in Fig. 2. Earlier studies have shown that Cal 99 probably evolved from Ark DPI [18].

Bottom Line: The genome of Ark DPI consists of 27,620 nucleotides, excluding poly (A) tail, and comprises ten open reading frames.Possible recombination sites were found at the intergenic region of spike gene, 3'end of S1 and 3a gene.Independent recombination events may have occurred in the entire genome of Ark DPI, involving four different IBV strains, suggesting that genomic RNA recombination may occur in any part of the genome at number of sites.

View Article: PubMed Central - HTML - PubMed

Affiliation: Center of Marine Biotechnology, University of Maryland Biotechnology Institute, Baltimore, 701 East Pratt Street, Baltimore, Maryland 21202-3101, USA. ammayapp@umbi.umd.edu

ABSTRACT
An infectious bronchitis virus Arkansas DPI (Ark DPI) virulent strain was sequenced, analyzed and compared with many different IBV strains and coronaviruses. The genome of Ark DPI consists of 27,620 nucleotides, excluding poly (A) tail, and comprises ten open reading frames. Comparative sequence analysis of Ark DPI with other IBV strains shows striking similarity to the Conn, Gray, JMK, and Ark 99, which were circulating during that time period. Furthermore, comparison of the Ark genome with other coronaviruses demonstrates a close relationship to turkey coronavirus. Among non-structural genes, the 5'untranslated region (UTR), 3C-like proteinase (3CLpro) and the polymerase (RdRp) sequences are 100% identical to the Gray strain. Among structural genes, S1 has 97% identity with Ark 99; S2 has 100% identity with JMK and 96% to Conn; 3b 99%, and 3C to N is 100% identical to Conn strain. Possible recombination sites were found at the intergenic region of spike gene, 3'end of S1 and 3a gene. Independent recombination events may have occurred in the entire genome of Ark DPI, involving four different IBV strains, suggesting that genomic RNA recombination may occur in any part of the genome at number of sites. Hence, we speculate that the Ark DPI strain originated from the Conn strain, but diverged and evolved independently by point mutations and recombination between field strains.

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Related in: MedlinePlus