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Nuclear accumulation of measles virus nucleoprotein associated with a temperature-sensitive mutant
Abstract:
A measles virus (MV) Lec strain conditional-lethal (temperature-sensitive) mutant, designated MV ts38, has been isolated from 5-fluorouracil-mutagenized stock. The mutant has been characterized with regard to growth characteristics at 32 degrees C (permissive temperature) and 39 degrees C (non-permissive temperature). Virus-specific RNA transcription and/or translation appeared to be blocked at the non-permissive temperature as no virus-specific products could be detected by biochemical or immunological procedures. Following initiation of viral replication at 32 degrees C, with subsequent shift-up to 39 degrees C, presynthesized nucleoprotein (NP) was transported to, and accumulated in the cell nucleus whereas other viral proteins could not be detected there. A corresponding accumulation of NP in the nucleus is most often seen in association with MV neurotropic subacute sclerosing panencephalitis isolates in vivo and in vitro.
Insights
A temperature-sensitive measles virus (MV) mutant, MV ts38, blocks viral RNA transcription and translation at non-permissive temperatures. Nucleoprotein accumulates in the nucleus, similar to neurotropic MV strains.
Area of Science:
- Virology
- Molecular Biology
- Genetics
Background:
- Measles virus (MV) can cause severe neurological diseases.
- Understanding MV replication is crucial for developing effective treatments.
- Conditional-lethal mutants are valuable tools for studying viral gene function.
Purpose of the Study:
- To characterize a novel temperature-sensitive mutant of measles virus, MV ts38.
- To investigate the molecular mechanisms underlying MV replication defects at restrictive temperatures.
- To explore the potential link between MV ts38's behavior and neurovirulence.
Main Methods:
- Isolation and characterization of MV ts38 from mutagenized stock.
- Growth studies at permissive (32°C) and non-permissive (39°C) temperatures.
- Biochemical and immunological assays to detect viral RNA and protein synthesis.
- Analysis of viral protein localization within infected cells.
Main Results:
- MV ts38 exhibited conditional-lethal growth, with replication blocked at 39°C.
- Viral RNA transcription and/or translation were inhibited at the non-permissive temperature.
- Upon temperature shift, nucleoprotein (NP) localized to the nucleus, while other viral proteins did not.
- This NP nuclear accumulation mirrors patterns observed in neurotropic MV isolates.
Conclusions:
- MV ts38 represents a valuable tool for studying measles virus gene function.
- The temperature-sensitive defect in MV ts38 likely affects viral RNA processing or translation.
- Nuclear accumulation of NP in MV ts38-infected cells suggests a potential link to MV neurotropism.