Mutation in murine coronavirus replication protein nsp4 alters assembly of double membrane vesicles
| Autor: | Susan C. Baker, Mark A. Clementz, Timothy E. O'Brien, Amornrat Kanjanahaluethai |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
animal structures
Glycosylation viruses RNA-dependent RNA polymerase Viral Nonstructural Proteins Biology medicine.disease_cause Article Virus Cell Line 03 medical and health sciences chemistry.chemical_compound Microscopy Electron Transmission Cricetinae Virology medicine Animals Humans Nonstructural proteins 030304 developmental biology Coronavirus Murine hepatitis virus 0303 health sciences 030306 microbiology Virus Assembly Cell Membrane RNA biochemical phenomena metabolism and nutrition Molecular biology Reverse genetics Mitochondria 3. Good health Double membrane vesicles Amino Acid Substitution chemistry Viral replication Cytoplasm Mutagenesis Site-Directed ts mutant |
| Zdroj: | Virology |
| ISSN: | 0042-6822 |
| DOI: | 10.1016/j.virol.2008.01.018 |
| Popis: | Coronaviruses are positive-strand RNA viruses that replicate in the cytoplasm of infected cells by generating a membrane-associated replicase complex. The replicase complex assembles on double membrane vesicles (DMVs). Here, we studied the role of a putative replicase anchor, nonstructural protein 4 (nsp4), in the assembly of murine coronavirus DMVs. We used reverse genetics to generate infectious clone viruses (icv) with an alanine substitution at nsp4 glycosylation site N176 or N237, or an asparagine to threonine substitution (nsp4-N258T), which is proposed to confer a temperature sensitive phenotype. We found that nsp4-N237A is lethal and nsp4-N258T generated a virus (designated Alb ts6 icv) that is temperature sensitive for viral replication. Analysis of Alb ts6 icv-infected cells revealed that there was a dramatic reduction in DMVs and that both nsp4 and nsp3 partially localized to mitochondria when cells were incubated at the non-permissive temperature. These results reveal a critical role of nsp4 in directing coronavirus DMV assembly. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect