Coronavirus pathogenesis and host response to infection

There are eight open reading frames (ORF’s) in the SARS-coronavirus (SARS-CoV) genome that encode non-structural proteins whose functions are currently unknown. I am currently investigating ORF6, which when expressed in the context of a recombinant mouse hepatitis virus, enhances virulence, converting a non-lethal infection into a uniformly lethal infection. My project aims to uncover the function of ORF6 by developing a reverse genetics system and animal model for SARS-CoV that will allow me to manipulate the genome and assay for changes in pathogenesis. In addition, I will be using biochemical techniques to characterize the ORF6 protein and determine its structure, as this may give insight into its function.

Netland J, DeDiego ML, Zhao J, Fett C, Alvarez E, Nieto-Torres JL, Enjuanes L, Perlman S. Immunization with an attenuated severe acute respiratory syndrome coronavirus deleted in E protein protects against lethal respiratory disease. Virology. 2010 Mar 30;399(1):120-8. Epub 2010 Jan 27. PubMed PMID: 20110095; PubMed Central PMCID: PMC2830353.

Zhou H, Ferraro D, Zhao J, Hussain S, Shao J, Trujillo J, Netland J, Gallagher T, Perlman S. The N-terminal region of severe acute respiratory syndrome coronavirus protein 6 induces membrane rearrangement and enhances virus replication. J Virol. 2010 Apr;84(7):3542-51. Epub 2010 Jan 27. PubMed PMID:20106914; PubMed Central PMCID: PMC2838104.

Perlman S, Netland J. Coronaviruses post-SARS: update on replication and pathogenesis. Nat Rev Microbiol. 2009 Jun;7(6):439-50. Review. PubMed PMID:19430490; PubMed Central PMCID: PMC2830095.

Zhao J, Falcón A, Zhou H, Netland J, Enjuanes L, Pérez Breña P, Perlman S. Severe acute respiratory syndrome coronavirus protein 6 is required for optimal replication. J Virol. 2009 Mar;83(5):2368-73. Epub 2008 Dec 17. PubMed PMID:19091867; PubMed Central PMCID: PMC2643704.

Netland J, Meyerholz DK, Moore S, Cassell M, Perlman S. Severe acute respiratory syndrome coronavirus infection causes neuronal death in the absence of encephalitis in mice transgenic for human ACE2. J Virol. 2008 Aug;82(15):7264-75. Epub 2008 May 21. PubMed PMID: 18495771; PubMed Central PMCID: PMC2493326.

Netland J, Ferraro D, Pewe L, Olivares H, Gallagher T, Perlman S. Enhancement of murine coronavirus replication by severe acute respiratory syndrome coronavirus protein 6 requires the N-terminal hydrophobic region but not C-terminal sorting motifs. J Virol. 2007 Oct;81(20):11520-5. Epub 2007 Aug 1. PubMed PMID: 17670827; PubMed Central PMCID: PMC2045524.

Tangudu C, Olivares H, Netland J, Perlman S, Gallagher T. Severe acute respiratory syndrome coronavirus protein 6 accelerates murine coronavirus infections. J Virol. 2007 Feb;81(3):1220-9. Epub 2006 Nov 15. PubMed PMID:17108045; PubMed Central PMCID: PMC1797517.

McCray PB Jr, Pewe L, Wohlford-Lenane C, Hickey M, Manzel L, Shi L, Netland J, Jia HP, Halabi C, Sigmund CD, Meyerholz DK, Kirby P, Look DC, Perlman S. Lethal infection of K18-hACE2 mice infected with severe acute respiratory syndrome coronavirus. J Virol. 2007 Jan;81(2):813-21. Epub 2006 Nov 1. PubMed PMID:17079315; PubMed Central PMCID: PMC1797474.

Pewe L, Zhou H, Netland J, Tangadu C, Olivares H, Shi L, Look D, Gallagher T, Perlman S. A SARS-CoV-specific protein enhances virulence of an attenuated strain of mouse hepatitis virus. Adv Exp Med Biol. 2006;581:493-8. PubMed PMID:17037583.

Jia HP, Look DC, Hickey M, Shi L, Pewe L, Netland J, Farzan M, Wohlford-Lenane C, Perlman S, McCray PB Jr. Infection of human airway epithelia by SARS coronavirus is associated with ACE2 expression and localization. Adv Exp Med Biol. 2006;581:479-84. Review. PubMed PMID: 17037581.

Jia HP, Look DC, Shi L, Hickey M, Pewe L, Netland J, Farzan M, Wohlford-Lenane C, Perlman S, McCray PB Jr. ACE2 receptor expression and severe acute respiratory syndrome coronavirus infection depend on differentiation of human airway epithelia. J Virol. 2005 Dec;79(23):14614-21. PubMed PMID: 16282461; PubMed Central PMCID: PMC1287568.

Pewe L, Zhou H, Netland J, Tangudu C, Olivares H, Shi L, Look D, Gallagher T, Perlman S. A severe acute respiratory syndrome-associated coronavirus-specific protein enhances virulence of an attenuated murine coronavirus. J Virol. 2005 Sep;79(17):11335-42. PubMed PMID: 16103185; PubMed Central PMCID: PMC1193615.

Pewe LL, Netland JM, Heard SB, Perlman S. Very diverse CD8 T cell clonotypic responses after virus infections. J Immunol. 2004 Mar 1;172(5):3151-6. PubMed PMID: 14978121.