Identification of two functional domains within the arenavirus nucleoprotein

Levingston Macleod, Jesica M., D'Antuono, Alejandra, Loureiro, Maria Eugenia, Casabona, Juan Cruz, Gomez, Guillermo A. and Lopez, Nora (2011) Identification of two functional domains within the arenavirus nucleoprotein. Journal of Virology, 85 5: 2012-2023. doi:10.1128/JVI.01875-10

Author Levingston Macleod, Jesica M.
D'Antuono, Alejandra
Loureiro, Maria Eugenia
Casabona, Juan Cruz
Gomez, Guillermo A.
Lopez, Nora
Title Identification of two functional domains within the arenavirus nucleoprotein
Journal name Journal of Virology   Check publisher's open access policy
ISSN 0022-538X
Publication date 2011-03
Year available 2010
Sub-type Article (original research)
DOI 10.1128/JVI.01875-10
Open Access Status DOI
Volume 85
Issue 5
Start page 2012
End page 2023
Total pages 12
Place of publication Washington, D.C. U.S.A.
Publisher American Society for Microbiology
Collection year 2012
Language eng
Abstract Tacaribe virus (TCRV) belongs to the Arenaviridae family. Its bisegmented negative-stranded RNA genome encodes the nucleoprotein (N), the precursor of the envelope glycoproteins, the polymerase (L), and a RING finger matrix (Z) protein. The 570-amino-acid N protein binds to viral RNA, forming nucleocapsids, which are the template for transcription and replication by the viral polymerase. We have previously shown that the interaction between N and Z is required for assembly of infectious virus-like particles (VLPs) (J. C. Casabona et al., J. Virol. 83:7029-7039, 2009). Here, we examine the functional organization of TCRV N protein. A series of deletions and point mutations were introduced into the N-coding sequence, and the ability of the mutants to sustain heterotypic (N-Z) or homotypic (N-N) interactions was analyzed. We found that N protein displays two functional domains. By using coimmunoprecipitation studies, VLP incorporation assays, and double immunofluorescence staining, the carboxy-terminal region of N was found to be required for N-Z interaction and also necessary for incorporation of N protein into VLPs. Moreover, further analysis of this region showed that the integrity of a putative zinc-finger motif, as well as its amino-flanking sequence (residues 461 to 489), are critical for Z binding and N incorporation into VLPs. In addition, we provide evidence of an essential role of the amino-terminal region of N protein for N-N interaction. In this regard, using reciprocal coimmunoprecipitation analysis, we identified a 28-residue region predicted to form a coiled-coil domain (residues 92 to 119) as a newly recognized molecular determinant of N homotypic interactions.
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ
Additional Notes Published Ahead of Print 15 December 2010.

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2012 Collection
ERA 2012 Admin Only
Institute for Molecular Bioscience - Publications
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Created: Thu, 20 Oct 2011, 08:57:23 EST by Guillermo Gomez on behalf of Institute for Molecular Bioscience