K48-linked polyubiquitination of dengue virus NS1 protein inhibits its interaction with the viral partner NS4B

Maria Giraldo Giraldo, Oscar Vargas-Cuartas, Juan Carlos Gallego-Gomez, Pei-Yong Shi, Leonardo Padilla-Sanabria, Jhon Carlos Castaño-Osorio, Ricardo Rajsbaum Gorodezky

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Dengue virus (DENV) is a member of the Flaviviridae family, which is transmitted to mammalian species through arthropods, and causes dengue fever or severe dengue fever in humans. The DENV genome encodes for multiple nonstructural (NS) proteins including NS1. NS1 plays an essential role in replication by interacting with other viral proteins including NS4B, however how these interactions are regulated during virus infection is not known. By using bioinformatics, mass spectrometry analysis, and co-immunoprecipitation assays, here we show that DENV-NS1 is ubiquitinated on multiples lysine residues during DENV infection, including K189, a lysine residue previously shown to be important for efficient DENV replication. Data from in vitro and cell culture experiments indicate that dengue NS1 undergoes modification with K48-linked polyubiquitin chains, which usually target proteins to the proteasome for degradation. Furthermore, ubiquitinated NS1 was detected in lysates as well as in supernatants of human and mosquito infected cells. Ubiquitin deconjugation of NS1 using the deubiquitinase OTU resulted in increased interaction with the viral protein NS4B suggesting that ubiquitinated NS1 has reduced affinity for NS4B. In support of these data, a K189R mutation on NS1, which abrogates ubiquitination on amino acid residue 189 of NS1, also increased NS1-NS4B interactions. Our work describes a new mechanism of regulation of NS1-NS4B interactions and suggests that ubiquitination of NS1 may affect DENV replication.

Original languageEnglish (US)
Pages (from-to)1-11
Number of pages11
JournalVirus Research
Volume246
DOIs
StatePublished - Feb 15 2018

Fingerprint

Dengue Virus
Dengue
Proteins
Ubiquitination
Viral Proteins
Virus Diseases
Virus Replication
Lysine
Flaviviridae
Polyubiquitin
Severe Dengue
Arthropods
Proteasome Endopeptidase Complex
Ubiquitin
Computational Biology
Culicidae
Immunoprecipitation
Mass Spectrometry
Cell Culture Techniques
Genome

Keywords

  • Dengue virus
  • Deubiquitinase
  • K48-linked polyubiquitin
  • NS1
  • NS4B
  • Proteasome
  • Ubiquitination

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases
  • Cancer Research

Cite this

K48-linked polyubiquitination of dengue virus NS1 protein inhibits its interaction with the viral partner NS4B. / Giraldo Giraldo, Maria; Vargas-Cuartas, Oscar; Gallego-Gomez, Juan Carlos; Shi, Pei-Yong; Padilla-Sanabria, Leonardo; Castaño-Osorio, Jhon Carlos; Rajsbaum Gorodezky, Ricardo.

In: Virus Research, Vol. 246, 15.02.2018, p. 1-11.

Research output: Contribution to journalArticle

Giraldo Giraldo, Maria ; Vargas-Cuartas, Oscar ; Gallego-Gomez, Juan Carlos ; Shi, Pei-Yong ; Padilla-Sanabria, Leonardo ; Castaño-Osorio, Jhon Carlos ; Rajsbaum Gorodezky, Ricardo. / K48-linked polyubiquitination of dengue virus NS1 protein inhibits its interaction with the viral partner NS4B. In: Virus Research. 2018 ; Vol. 246. pp. 1-11.
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abstract = "Dengue virus (DENV) is a member of the Flaviviridae family, which is transmitted to mammalian species through arthropods, and causes dengue fever or severe dengue fever in humans. The DENV genome encodes for multiple nonstructural (NS) proteins including NS1. NS1 plays an essential role in replication by interacting with other viral proteins including NS4B, however how these interactions are regulated during virus infection is not known. By using bioinformatics, mass spectrometry analysis, and co-immunoprecipitation assays, here we show that DENV-NS1 is ubiquitinated on multiples lysine residues during DENV infection, including K189, a lysine residue previously shown to be important for efficient DENV replication. Data from in vitro and cell culture experiments indicate that dengue NS1 undergoes modification with K48-linked polyubiquitin chains, which usually target proteins to the proteasome for degradation. Furthermore, ubiquitinated NS1 was detected in lysates as well as in supernatants of human and mosquito infected cells. Ubiquitin deconjugation of NS1 using the deubiquitinase OTU resulted in increased interaction with the viral protein NS4B suggesting that ubiquitinated NS1 has reduced affinity for NS4B. In support of these data, a K189R mutation on NS1, which abrogates ubiquitination on amino acid residue 189 of NS1, also increased NS1-NS4B interactions. Our work describes a new mechanism of regulation of NS1-NS4B interactions and suggests that ubiquitination of NS1 may affect DENV replication.",
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