Abstract
Hantaviruses (Bunyaviridae) are negative-strand RNA viruses with a tripartite genome. The small (S) segment encodes the nucleocapsid protein and, in some hantaviruses, also the nonstructural protein (NSs). The aim of this study was to find potential cellular partners for the hantaviral NSs protein. Toward this aim, yeast two-hybrid (Y2H) screening of mouse cDNA library was performed followed by a search for potential NSs protein counterparts via analyzing a cellular interactome. The resulting interaction network was shown to form logical, clustered structures. Furthermore, several potential binding partners for the NSs protein, for instance ACBD3, were identified and, to prove the principle, interaction between NSs and ACBD3 proteins was demonstrated biochemically.
Highlights
Hantaviruses constitute the Hantavirus genus in the family Bunyaviridae [1]
Using Tula virus (TULV) and Puumala virus (PUUV) as models it was shown that the NSs open reading frame (ORF) is functional and the product interferes with interferon (IFN) production by inhibiting the activity of IFN-b promoter and NF-kB- and IRF-3-responsive promoters [6]
To verify that NSs proteins are expressed from yeasts transformed with their respectives plasmids, the colonies from SD/-Leu/-Trp plates containing the following samples: (1) empty pGBKT7 - empty pACT2, (2) pGBKT7-PUUV NSs - empty pACT2, and (3) pGBKT7-TULV NSs - empty pACT2 were transferred to SD/-Leu/-Trp liquid medium and incubated at +30uC on a shaker overnight
Summary
Hantaviruses constitute the Hantavirus genus in the family Bunyaviridae [1]. Natural hosts for hantaviruses are mostly Muroidea rodents of Muridae and Cricetidae families (for the current rodent taxonomy, see [2]) and insectivores of family Soricidae. Hantaviruses are negative-strand RNA viruses (NSRV) with a tripartite genome consisting of Large (L), Medium (M) and Small (S) segments [1,4,5]. The L segment encodes the viral RNAdependent RNA polymerase (L protein). In some hantaviruses the S segment has an open reading frame (ORF) for a nonstructural protein (NSs) [5]. Using Tula virus (TULV) and Puumala virus (PUUV) as models it was shown that the NSs ORF is functional and the product interferes with interferon (IFN) production by inhibiting the activity of IFN-b promoter and NF-kB- and IRF-3-responsive promoters [6]. Functions of the hantaviral NSs protein other than IFN inhibition (e.g., regulation of viral transcription/replication or RNA-binding) remain unknown
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