Characterization of Interaction between Viral Protein Nsp1 and Cellular Proteins in SARS Coronavirus

Author(s)

Maya DodhiaFollow

Department, Center, or Institute

Biology

Presentation Format

Poster

Presentation Type

On-campus research

Description

Severe acute respiratory syndrome (SARS) is a contagious and potentially fatal respiratory illness that is transmitted via airborne respiratory droplets and human contact with other infected individuals. SARS is caused by a coronavirus (CoV), which are common viruses responsible for causing a variety of respiratory infections in humans and animals. According to the Centers for Disease Control and the World Health Organization, this viral respiratory disease first appeared in China in 2002, leading to an worldwide epidemic that resulted in more than 8,000 cases. Ten percent of these cases resulted in deaths within the span of a year in 26 different countries. SARS-CoV produces sixteen nonstructural proteins including nonstructural protein 1 (nsp1), which is the key viral protein implicated in the down-regulation of host gene expression. In this study, we detected a mechanism by which nsp1 alters the composition of the nuclear pore complex and subsequently disrupts nuclear-cytoplasmic translocation of key RNA binding proteins. Using GST pull-down followed by LC-MS/MS, we identified several cellular proteins that are associated with nsp1. Proteins isolated by mass spectrometry were independently verified by anti-Myc immunoprecipitation followed by western blot analyses. Using indirect immunofluorescence, we showed that expression of nsp1 in HEK cells disrupts Nup93 localization, whereby Nup93 displaces from the nuclear envelope and translocates to the nucleoplasm. These results indicate that nsp1 alters host gene expression through the disruption of the nuclear pore complex and alteration of nuclear-cytoplasmic transport of biomolecules.

Session Number

4

Start Date and Time

4-9-2019 3:00 PM

Location

PAC Gym

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Apr 9th, 3:00 PM

Characterization of Interaction between Viral Protein Nsp1 and Cellular Proteins in SARS Coronavirus

PAC Gym

Severe acute respiratory syndrome (SARS) is a contagious and potentially fatal respiratory illness that is transmitted via airborne respiratory droplets and human contact with other infected individuals. SARS is caused by a coronavirus (CoV), which are common viruses responsible for causing a variety of respiratory infections in humans and animals. According to the Centers for Disease Control and the World Health Organization, this viral respiratory disease first appeared in China in 2002, leading to an worldwide epidemic that resulted in more than 8,000 cases. Ten percent of these cases resulted in deaths within the span of a year in 26 different countries. SARS-CoV produces sixteen nonstructural proteins including nonstructural protein 1 (nsp1), which is the key viral protein implicated in the down-regulation of host gene expression. In this study, we detected a mechanism by which nsp1 alters the composition of the nuclear pore complex and subsequently disrupts nuclear-cytoplasmic translocation of key RNA binding proteins. Using GST pull-down followed by LC-MS/MS, we identified several cellular proteins that are associated with nsp1. Proteins isolated by mass spectrometry were independently verified by anti-Myc immunoprecipitation followed by western blot analyses. Using indirect immunofluorescence, we showed that expression of nsp1 in HEK cells disrupts Nup93 localization, whereby Nup93 displaces from the nuclear envelope and translocates to the nucleoplasm. These results indicate that nsp1 alters host gene expression through the disruption of the nuclear pore complex and alteration of nuclear-cytoplasmic transport of biomolecules.