SARS coronavirus nsp1 protein induces template-dependent endonucleolytic cleavage of mRNAs: Viral mRNAs are resistant to nsp1-induced RNA cleavage

Cheng Huang, Kumari G. Lokugamage, Janet M. Rozovics, Krishna Narayanan, Bert L. Semler, Shinji Makino

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

SARS coronavirus (SCoV) nonstructural protein (nsp) 1, a potent inhibitor of host gene expression, possesses a unique mode of action: it binds to 40S ribosomes to inactivate their translation functions and induces host mRNA degradation. Our previous study demonstrated that nsp1 induces RNA modification near the 5′-end of a reporter mRNA having a short 5′ untranslated region and RNA cleavage in the encephalomyocarditis virus internal ribosome entry site (IRES) region of a dicistronic RNA template, but not in those IRES elements from hepatitis C or cricket paralysis viruses. By using primarily cell-free, in vitro translation systems, the present study revealed that the nsp1 induced endonucleolytic RNA cleavage mainly near the 5′ untranslated region of capped mRNA templates. Experiments using dicistronic mRNAs carrying different IRESes showed that nsp1 induced endonucleolytic RNA cleavage within the ribosome loading region of type I and type II picornavirus IRES elements, but not that of classical swine fever virus IRES, which is characterized as a hepatitis C virus-like IRES. The nsp1-induced RNA cleavage of template mRNAs exhibited no apparent preference for a specific nucleotide sequence at the RNA cleavage sites. Remarkably, SCoV mRNAs, which have a 5′ cap structure and 3′ poly A tail like those of typical host mRNAs, were not susceptible to nsp1-mediated RNA cleavage and importantly, the presence of the 5′-end leader sequence protected the SCoV mRNAs from nsp1-induced endonucleolytic RNA cleavage. The escape of viral mRNAs from nsp1-induced RNA cleavage may be an important strategy by which the virus circumvents the action of nsp1 leading to the efficient accumulation of viral mRNAs and viral proteins during infection.

Original languageEnglish (US)
Article numbere1002433
JournalPLoS Pathogens
Volume7
Issue number12
DOIs
StatePublished - Dec 2011

Fingerprint

RNA Cleavage
Messenger RNA
SARS Virus
5' Untranslated Regions
Ribosomes
SARS coronavirus Nsp1 protein
Classical swine fever virus
RNA
Encephalomyocarditis virus
Viruses
Picornaviridae
Gryllidae
Untranslated RNA
RNA Stability
Viral Proteins
Hepatitis C
Paralysis
Hepacivirus
Internal Ribosome Entry Sites

ASJC Scopus subject areas

  • Microbiology
  • Parasitology
  • Virology
  • Immunology
  • Genetics
  • Molecular Biology

Cite this

SARS coronavirus nsp1 protein induces template-dependent endonucleolytic cleavage of mRNAs : Viral mRNAs are resistant to nsp1-induced RNA cleavage. / Huang, Cheng; Lokugamage, Kumari G.; Rozovics, Janet M.; Narayanan, Krishna; Semler, Bert L.; Makino, Shinji.

In: PLoS Pathogens, Vol. 7, No. 12, e1002433, 12.2011.

Research output: Contribution to journalArticle

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