Evidence for ubiquitin-regulated nuclear and subnuclear trafficking among Paramyxovirinae matrix proteins

Mickey Pentecost, Ajay A. Vashisht, Talia Lester, Tim Voros, Shannon M. Beaty, Arnold Park, Yao E. Wang, Tatyana E. Yun, Alexander Freiberg, James A. Wohlschlegel, Benhur Lee

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

The paramyxovirus matrix (M) protein is a molecular scaffold required for viral morphogenesis and budding at the plasma membrane. Transient nuclear residence of some M proteins hints at non-structural roles. However, little is known regarding the mechanisms that regulate the nuclear sojourn. Previously, we found that the nuclear-cytoplasmic trafficking of Nipah virus M (NiV-M) is a prerequisite for budding, and is regulated by a bipartite nuclear localization signal (NLSbp), a leucine-rich nuclear export signal (NES), and monoubiquitination of the K258 residue within the NLSbp itself (NLSbp-lysine). To define whether the sequence determinants of nuclear trafficking identified in NiV-M are common among other Paramyxovirinae M proteins, we generated the homologous NES and NLSbp-lysine mutations in M proteins from the five major Paramyxovirinae genera. Using quantitative 3D confocal microscopy, we determined that the NES and NLSbp-lysine are required for the efficient nuclear export of the M proteins of Nipah virus, Hendra virus, Sendai virus, and Mumps virus. Pharmacological depletion of free ubiquitin or mutation of the conserved NLSbp-lysine to an arginine, which inhibits M ubiquitination, also results in nuclear and nucleolar retention of these M proteins. Recombinant Sendai virus (rSeV-eGFP) bearing the NES or NLSbp-lysine M mutants rescued at similar efficiencies to wild type. However, foci of cells expressing the M mutants displayed marked fusogenicity in contrast to wild type, and infection did not spread. Recombinant Mumps virus (rMuV-eGFP) bearing the homologous mutations showed similar defects in viral morphogenesis. Finally, shotgun proteomics experiments indicated that the interactomes of Paramyxovirinae M proteins are significantly enriched for components of the nuclear pore complex, nuclear transport receptors, and nucleolar proteins. We then synthesize our functional and proteomics data to propose a working model for the ubiquitin-regulated nuclear-cytoplasmic trafficking of cognate paramyxovirus M proteins that show a consistent nuclear trafficking phenotype.

Original languageEnglish (US)
Pages (from-to)e1004739
JournalPLoS Pathogens
Volume11
Issue number3
DOIs
StatePublished - Mar 1 2015

Fingerprint

Paramyxovirinae
Ubiquitin
Nuclear Export Signals
Nipah Virus
Lysine
Proteins
Mumps virus
Sendai virus
Cell Nucleus Active Transport
Nuclear Proteins
Morphogenesis
Proteomics
Mutation
Hendra Virus
Nuclear Pore
Nuclear Localization Signals
Ubiquitination
Firearms
Cytoplasmic and Nuclear Receptors
Confocal Microscopy

ASJC Scopus subject areas

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

Cite this

Pentecost, M., Vashisht, A. A., Lester, T., Voros, T., Beaty, S. M., Park, A., ... Lee, B. (2015). Evidence for ubiquitin-regulated nuclear and subnuclear trafficking among Paramyxovirinae matrix proteins. PLoS Pathogens, 11(3), e1004739. https://doi.org/10.1371/journal.ppat.1004739

Evidence for ubiquitin-regulated nuclear and subnuclear trafficking among Paramyxovirinae matrix proteins. / Pentecost, Mickey; Vashisht, Ajay A.; Lester, Talia; Voros, Tim; Beaty, Shannon M.; Park, Arnold; Wang, Yao E.; Yun, Tatyana E.; Freiberg, Alexander; Wohlschlegel, James A.; Lee, Benhur.

In: PLoS Pathogens, Vol. 11, No. 3, 01.03.2015, p. e1004739.

Research output: Contribution to journalArticle

Pentecost, M, Vashisht, AA, Lester, T, Voros, T, Beaty, SM, Park, A, Wang, YE, Yun, TE, Freiberg, A, Wohlschlegel, JA & Lee, B 2015, 'Evidence for ubiquitin-regulated nuclear and subnuclear trafficking among Paramyxovirinae matrix proteins', PLoS Pathogens, vol. 11, no. 3, pp. e1004739. https://doi.org/10.1371/journal.ppat.1004739
Pentecost, Mickey ; Vashisht, Ajay A. ; Lester, Talia ; Voros, Tim ; Beaty, Shannon M. ; Park, Arnold ; Wang, Yao E. ; Yun, Tatyana E. ; Freiberg, Alexander ; Wohlschlegel, James A. ; Lee, Benhur. / Evidence for ubiquitin-regulated nuclear and subnuclear trafficking among Paramyxovirinae matrix proteins. In: PLoS Pathogens. 2015 ; Vol. 11, No. 3. pp. e1004739.
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