Analysis of SARS-CoV-2 nucleocapsid protein sequence variations in ASEAN countries
DOI:
https://doi.org/10.13181/mji.oa.215304Keywords:
nucleocapsid proteins, phylogeny, SARS-CoV-2, sequence alignmentAbstract
BACKGROUND Nucleocapsid (N) protein is one of four structural proteins of SARS-CoV-2 which is known to be more conserved than spike protein and is highly immunogenic. This study aimed to analyze the variation of the SARS-CoV-2 N protein sequences in ASEAN countries, including Indonesia.
METHODS Complete sequences of SARS-CoV-2 N protein from each ASEAN country were obtained from Global Initiative on Sharing All Influenza Data (GISAID), while the reference sequence was obtained from GenBank. All sequences collected from December 2019 to March 2021 were grouped to the clade according to GISAID, and two representative isolates were chosen from each clade for the analysis. The sequences were aligned by MUSCLE, and phylogenetic trees were built using MEGA-X software based on the nucleotide and translated AA sequences.
RESULTS 98 isolates of complete N protein genes from ASEAN countries were analyzed. The nucleotides of all isolates were 97.5% conserved. Of 31 nucleotide changes, 22 led to amino acid (AA) substitutions; thus, the AA sequences were 94.5% conserved. The phylogenetic tree of nucleotide and AA sequences shows similar branches. Nucleotide variations in clade O (C28311T); clade GR (28881–28883 GGG>AAC); and clade GRY (28881–28883 GGG>AAC and C28977T) lead to specific branches corresponding to the clade within both trees.
CONCLUSIONS The N protein sequences of SARS-CoV-2 across ASEAN countries are highly conserved. Most isolates were closely related to the reference sequence originating from China, except the isolates representing clade O, GR, and GRY which formed specific branches in the phylogenetic tree.
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References
Wang Z, Qiang W, Ke H. A handbook of 2019-nCoV pneumonia control and prevention. Hubei Sci Technol Press. 2020;1-108.
Peeri NC, Shrestha N, Rahman MS, Zaki R, Tan Z, Bibi S, et al. The SARS, MERS and novel coronavirus (COVID-19) epidemics, the newest and biggest global health threats: what lessons have we learned? Int J Epidemiol. 2020;49(3):717-26. https://doi.org/10.1093/ije/dyaa033
Suprobowati OD, Kurniati I. Virology Medical Laboratory Technology (TLM) Teaching Materials [Internet]. Jakarta: Ministry of Health of the Republic of Indonesia; 2018. [cited 2020 Jun 24]. Available from: http://perpus.poltekeskupang.ac.id/index.php?p=show_detail&id=3150&keywords=TLM. Indonesian.
Harapan H, Itoh N, Yufika A, Winardi W, Keam S, Te H, et al. Coronavirus disease 2019 (COVID-19): a literature review. J Infect Public Health. 2020;13(5):667-73. https://doi.org/10.1016/j.jiph.2020.03.019
Helmy YA, Fawzy M, Elaswad A, Sobieh A, Kenney SP, Shehata AA. The COVID-19 pandemic: a comprehensive review of taxonomy, genetics, epidemiology, diagnosis, treatment, and control. J Clin Med. 2020;9(4):1225. https://doi.org/10.3390/jcm9041225
Kopecky-Bromberg SA, Martínez-Sobrido L, Frieman M, Baric RA, Palese P. Severe acute respiratory syndrome coronavirus open reading frame (ORF) 3b, ORF 6, and nucleocapsid proteins function as interferon antagonists. J Virol. 2007;81(2):548-57. https://doi.org/10.1128/JVI.01782-06
Surjit M, Liu B, Kumar P, Chow VT, Lal SK. The nucleocapsid protein of the SARS coronavirus is capable of self-association through a C-terminal 209 amino acid interaction domain. Biochem Biophys Res Commun. 2004;317(4):1030-6. https://doi.org/10.1016/j.bbrc.2004.03.154
Shahhosseini N, Wong G, Kobinger GP, Chinikar S. SARS-CoV-2 spillover transmission due to recombination event. Gene Rep. 2021;23:101045. https://doi.org/10.1016/j.genrep.2021.101045
Zeng W, Liu G, Ma H, Zhao D, Yang Y, Liu M, et al. Biochemical characterization of SARS-CoV-2 nucleocapsid protein. Biochem Biophys Res Commun. 2020;527(3):618-23. https://doi.org/10.1016/j.bbrc.2020.04.136
Dutta NK, Mazumdar K, Gordy JT. The nucleocapsid protein of SARS-CoV-2: a target for vaccine development. J Virol. 2020;94(13):e00647-20. https://doi.org/10.1128/JVI.00647-20
Chen Y, Liu Q, Guo D. Emerging coronaviruses: genome structure, replication, and pathogenesis. J Med Virol. 2020;92(4):418-23. https://doi.org/10.1002/jmv.25681
Kang S, Yang M, Hong Z, Zhang L, Huang Z, Chen X, et al. Crystal structure of SARS-CoV-2 nucleocapsid protein RNA binding domain reveals potential unique drug targeting sites. Acta Pharm Sin B. 2020;10(7):1228-38. https://doi.org/10.1016/j.apsb.2020.04.009
Yong SK, Su PC, Yang YS. Molecular targets for the testing of COVID-19. Biotechnol J. 2020;15(6):e2000152. https://doi.org/10.1002/biot.202000152
Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol. 2018;35(6):1547-9. https://doi.org/10.1093/molbev/msy096
Azad S. Amino acids: its types and uses. Int J Clin Diagnostic Pathol. 2018;1(1):13-6.
Islam MR, Hoque MN, Rahman MS, Alam ASMRU, Akther M, Puspo JA, et al. Genome-wide analysis of SARS-CoV-2 virus strains circulating worldwide implicates heterogeneity. Sci Rep. 2020;10:1404. https://doi.org/10.1038/s41598-020-70812-6
Rahman MS, Islam MR, Alam ASMRU, Islam I, Hoque MN, Akter S, et al. Evolutionary dynamics of SARS-CoV-2 nucleocapsid protein and its consequences. J Med Virol. 2021;93(4):2177-95. https://doi.org/10.1002/jmv.26626
Arena F, Pollini S, Rossolini GM, Margaglione M. Summary of the available molecular methods for detection of SARS-CoV-2 during the ongoing pandemic. Int J Mol Sci. 2021;22(3):1298. https://doi.org/10.3390/ijms22031298
Khan KA, Cheung P. Presence of mismatches between diagnostic PCR assays and coronavirus SARS-CoV-2 genome. R Soc Open Sci. 2020;7(6):200636. https://doi.org/10.1098/rsos.200636
Guan Q, Sadykov M, Mfarrej S, Hala S, Naeem R, Nugmanova R, et al. A genetic barcode of SARS-CoV-2 for monitoring global distribution of different clades during the COVID-19 pandemic. Int J Infect Dis. 2020;100:216-23. https://doi.org/10.1016/j.ijid.2020.08.052
Colman PM. Effects of amino acid sequence changes on antibody-antigen interactions. Res Immunol. 1994;145(1):33-6. https://doi.org/10.1016/S0923-2494(94)80039-1
World Health Organization. Antigen-detection in the diagnosis of SARS-CoV-2 infection using rapid immunoassays: interim guidance, 11 September 2020. World Health Organization; 2020. Available from: https://apps.who.int/iris/handle/10665/334253.
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