MKRN1 induces degradation of West Nile virus capsid protein by functioning as an E3 ligase

Aram Ko, Eun Woo Lee, Jung Yong Yeh, Mi Ran Yang, Wonkyung Oh, Jin San Moon, Jaewhan Song

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

West Nile virus capsid protein (WNVCp) displays pathogenic toxicity via the apoptotic pathway. However, a cellular mechanism protective against this toxic effect has not been observed so far. Here, we identified Makorin ring finger protein 1 (MKRN1) as a novel E3 ubiquitin ligase for WNVCp. The cytotoxic effects of WNVCp as well as its expression levels were inhibited in U2OS cells that stably expressed MKRN1. Immunoprecipitation analyses revealed an interaction between MKRN1 and WNVCp. Domain analysis indicated that the C terminus of MKRN1 and the N terminus of WNVCp were required for the interaction. MKRN1 could induce WNVCp ubiquitination and degradation in a proteasome-dependent manner. Interestingly, the WNVCp mutant with amino acids 1 to 105 deleted WNVCp was degraded by MKRN1, whereas the mutant with amino acids 1 to 90 deleted was not. When three lysine sites at positions 101, 103, and 104 of WNVCp were replaced with alanine, MKRN1-mediated ubiquitination and degradation of the mutant were significantly inhibited, suggesting that these sites are required for the ubiquitination. Finally, U2OS cell lines stably expressing MKRN1 were resistant to cytotoxic effects of WNV. In contrast, cells depleted of MKRN1 were more susceptible to WNVCp cytotoxicity. Confirming this, overexpression of MKRN1 significantly reduced, but depletion of MKRN1 increased, WNV proliferation in 293T cells. Taken together, our results suggest that MKRN1 can protect cells from WNV by inducing WNVCp degradation.

Original languageEnglish
Pages (from-to)426-436
Number of pages11
JournalJournal of Virology
Volume84
Issue number1
DOIs
Publication statusPublished - 2010 Jan 1

Fingerprint

West Nile virus
Ubiquitin-Protein Ligases
Capsid Proteins
coat proteins
ligases
degradation
proteins
Ubiquitination
cytotoxicity
mutants
Proteolysis
Makorin ring finger protein 1
Amino Acids
ubiquitin-protein ligase
amino acids
HEK293 Cells
Poisons
proteasome endopeptidase complex
cells
Proteasome Endopeptidase Complex

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Ko, Aram ; Lee, Eun Woo ; Yeh, Jung Yong ; Yang, Mi Ran ; Oh, Wonkyung ; Moon, Jin San ; Song, Jaewhan. / MKRN1 induces degradation of West Nile virus capsid protein by functioning as an E3 ligase. In: Journal of Virology. 2010 ; Vol. 84, No. 1. pp. 426-436.
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abstract = "West Nile virus capsid protein (WNVCp) displays pathogenic toxicity via the apoptotic pathway. However, a cellular mechanism protective against this toxic effect has not been observed so far. Here, we identified Makorin ring finger protein 1 (MKRN1) as a novel E3 ubiquitin ligase for WNVCp. The cytotoxic effects of WNVCp as well as its expression levels were inhibited in U2OS cells that stably expressed MKRN1. Immunoprecipitation analyses revealed an interaction between MKRN1 and WNVCp. Domain analysis indicated that the C terminus of MKRN1 and the N terminus of WNVCp were required for the interaction. MKRN1 could induce WNVCp ubiquitination and degradation in a proteasome-dependent manner. Interestingly, the WNVCp mutant with amino acids 1 to 105 deleted WNVCp was degraded by MKRN1, whereas the mutant with amino acids 1 to 90 deleted was not. When three lysine sites at positions 101, 103, and 104 of WNVCp were replaced with alanine, MKRN1-mediated ubiquitination and degradation of the mutant were significantly inhibited, suggesting that these sites are required for the ubiquitination. Finally, U2OS cell lines stably expressing MKRN1 were resistant to cytotoxic effects of WNV. In contrast, cells depleted of MKRN1 were more susceptible to WNVCp cytotoxicity. Confirming this, overexpression of MKRN1 significantly reduced, but depletion of MKRN1 increased, WNV proliferation in 293T cells. Taken together, our results suggest that MKRN1 can protect cells from WNV by inducing WNVCp degradation.",
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MKRN1 induces degradation of West Nile virus capsid protein by functioning as an E3 ligase. / Ko, Aram; Lee, Eun Woo; Yeh, Jung Yong; Yang, Mi Ran; Oh, Wonkyung; Moon, Jin San; Song, Jaewhan.

In: Journal of Virology, Vol. 84, No. 1, 01.01.2010, p. 426-436.

Research output: Contribution to journalArticle

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