Vimentin interacts with heterogeneous nuclear ribonucleoproteins and dengue nonstructural protein 1 and is important for viral replication and release.
Our previous study using expression proteomics demonstrated that many proteins, particularly five forms of heterogeneous nuclear ribonucleoproteins (hnRNPs), were up-regulated in human endothelial cells upon dengue virus infection. To address functional significance of these proteins in response to dengue virus infection, we performed a functional proteomics study to identify hnRNPs-interacting proteins in the infected EA.hy926 cells. Immunoprecipitation followed by 2-D PAGE and mass spectrometric analyses revealed 18 and 13 interacting partners of hnRNP C1/C2 and hnRNP K, respectively. Interestingly, vimentin was a common partner for both hnRNP C1/C2 and K. The interaction between vimentin and these hnRNPs was confirmed by reciprocal immunoprecipitation followed by Western blot analysis and also by double immunofluorescence staining. Disruption of vimentin intermediate filament by acrylamide not only dissociated these complexes but also reduced nuclear hnRNPs expression, whereas cytosolic hnRNPs expression was unchanged. We also demonstrated that vimentin was strongly associated with dengue non-structural protein 1 (NS1). Disruption of vimentin intermediate filament not only dissociated this complex but also reduced dengue NS1 expression, as well as viral replication and release. Our data report for the first time that vimentin interacts with hnRNPs and dengue NS1, and plays a crucial role in replication and release of dengue virus.
Kanlaya
Rattiyaporn
R
Medical Proteomics Unit, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, 12th Floor Adulyadejvikrom Building, 2 Prannok Road, Bangkoknoi, Bangkok 10700, Thailand.
Pattanakitsakul
Sa-nga
SN
Sinchaikul
Supachok
S
Chen
Shui-Tein
ST
Thongboonkerd
Visith
V
D002460:Cell Line; D003716:Dengue Virus / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D034441:Heterogeneous-Nuclear Ribonucleoproteins / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D011485:Protein Binding; D053719:Tandem Mass Spectrometry; D014746:Vimentin / Q000378:metabolism*; D017361:Viral Nonstructural Proteins / Q000378:metabolism*
D034441:Heterogeneous-Nuclear Ribonucleoproteins; D014746:Vimentin; D017361:Viral Nonstructural Proteins