Identification and functional analysis of the SARS-COV-2 nucleocapsid protein

• Published in: BMC microbiology Vol. 21; no. 1; pp. 58 - 10
• Main Authors: Gao, Tianyi, Gao, Yingdong, Liu, Xiangxiang, Nie, Zhenlin, Sun, Huilin, Lin, Kang, Peng, Hongxin, Wang, Shukui
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 22.02.2021; BioMed Central; BMC
• Subjects: Amino Acid Sequence; Amino acids; Biological properties; Cell cycle; Coils; Coronaviridae; Coronaviruses; COVID-19; COVID-19 - genetics; COVID-19 - immunology; COVID-19 - virology; Epidemics; Flow cytometry; Functional analysis; Genome, Viral - genetics; Genomes; HCT116 Cells; Heat resistance; Humans; Hydrophobicity; Identification and classification; Kinases; Localization; Mass spectrometry; Mass spectroscopy; Middle East respiratory syndrome; Molecular Sequence Data; N protein; Nucleic acids; Nucleocapsid protein (N protein); Nucleocapsid Proteins - chemistry; Nucleocapsid Proteins - genetics; Nucleocapsid Proteins - metabolism; Nucleocapsids; Phosphorylation; Physicochemical properties; Physiological aspects; Protein kinase; Protein structure; Proteins; Random coil; Reproducibility of Results; S phase; SARS-COV-2; SARS-CoV-2 - genetics; SARS-CoV-2 - metabolism; SARS-CoV-2 - pathogenicity; Severe acute respiratory syndrome coronavirus 2; Structure; Target recognition; Tertiary structure; Therapeutic targets; Vaccines; Viral diseases; Viral proteins; Viral Vaccines - therapeutic use; Viruses; China; Phosphorylation; Structure; SARS-COV-2; Nucleocapsid protein (N protein)
• ISSN: 1471-2180; 1471-2180
• DOI: 10.1186/s12866-021-02107-3

A severe form of pneumonia, named coronavirus disease 2019 (COVID-19) by the World Health Organization is widespread on the whole world. The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was proved to be the main agent of COVID-19. In the present study, we conducted an in depth analysis of the SARS-COV-2 nucleocapsid to identify potential targets that may allow identification of therapeutic targets. The SARS-COV-2 N protein subcellular localization and physicochemical property was analyzed by PSORT II Prediction and ProtParam tool. Then SOPMA tool and swiss-model was applied to analyze the structure of N protein. Next, the biological function was explored by mass spectrometry analysis and flow cytometry. At last, its potential phosphorylation sites were analyzed by NetPhos3.1 Server and PROVEAN PROTEIN. SARS-COV-2 N protein composed of 419 aa, is a 45.6 kDa positively charged unstable hydrophobic protein. It has 91 and 49% similarity to SARS-CoV and MERS-CoV and is predicted to be predominantly a nuclear protein. It mainly contains random coil (55.13%) of which the tertiary structure was further determined with high reliability (95.76%). Cells transfected with SARS-COV-2 N protein usually show a G1/S phase block company with an increased expression of TUBA1C, TUBB6. At last, our analysis of SARS-COV-2 N protein predicted a total number of 12 phosphorylated sites and 9 potential protein kinases which would significantly affect SARS-COV-2 N protein function. In this study, we report the physicochemical properties, subcellular localization, and biological function of SARS-COV-2 N protein. The 12 phosphorylated sites and 9 potential protein kinase sites in SARS-COV-2 N protein may serve as promising targets for drug discovery and development for of a recombinant virus vaccine.