title: Dengue NS3, an RNAi suppressor, modulates the human miRNA pathways through its interacting partner.
abstract: RNAi acts as a host immune response against non-self molecules, including viruses. Viruses evolved to neutralize this response by expressing suppressor proteins. In the present study, we investigated dengue virus non structural protein 3 (dvNS3), for its RNAi-suppressor activity in human cell lines. Dengue virus (DV) NS3 reverts the GFP expression in GFP-silenced cell lines. Pull-down assays of dvNS3 revealed that it interacts with the host factor human heat shock cognate 70 (hHSC70). Down-regulation of hHSC70 resulted in accumulation of dengue viral genomic RNA. Also, the interaction of dvNS3 with hHSC70 perturbs the formation of RISC (RNA-induced silencing complex)-loading complex (RLC), by displacing TRBP (TAR RNA-binding protein) and possibly impairing the downstream activity of miRNAs. Interestingly, some of these miRNAs have earlier been reported to be down-regulated upon DV infection in Huh7 cells. Further studies on the miRNA-mRNA relationship along with mRNA profiling of samples overexpressing dvNS3 revealed up-regulation of TAZ (tafazzin) and SYNGR1 (synaptogyrin 1), known dengue viral host factors (DVHFs). Importantly, overexpression of dvNS3 in human embryonic kidney (HEK) 293T cells resulted in modulation of both mature and precursor miRNAs in human cell lines. Subsequent analysis suggested that dvNS3 induced stage-specific down-regulation of miRNAs. Taken together, these results suggest that dvNS3 affects biogenesis and function of host miRNAs to regulate DVHFs for favouring DV replication.
authors.affiliation: International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Pavan Kumar
authors.initials: PK
authors.lastname: Kakumani
authors.affiliation: International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: K S
authors.initials: KS
authors.lastname: Rajgokul
authors.affiliation: National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Sanket Singh
authors.initials: SS
authors.lastname: Ponia
authors.affiliation: International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Inderjeet
authors.initials: I
authors.lastname: Kaur
authors.affiliation: International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Srikrishna
authors.initials: S
authors.lastname: Mahanty
authors.affiliation: Translational Health Science and Technology Institute, Plot # 496, Phase-III, Udyog Vihar, Gurgaon, Haryana 122016, India.
authors.forename: Guruprasad R
authors.initials: GR
authors.lastname: Medigeshi
authors.affiliation: National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Akhil C
authors.initials: AC
authors.lastname: Banerjea
authors.affiliation: Department of Biotechnology, Hindustan College of Science and Technology, Farah, Mathura, Uttar Pradesh 281122, India.
authors.forename: Arun Prasad
authors.initials: AP
authors.lastname: Chopra
authors.affiliation: International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India pawanmal@gmail.com sunilm@icgeb.res.in raj@icgeb.res.in.
authors.forename: Pawan
authors.initials: P
authors.lastname: Malhotra
authors.affiliation: International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India pawanmal@gmail.com sunilm@icgeb.res.in raj@icgeb.res.in.
authors.forename: Sunil K
authors.initials: SK
authors.lastname: Mukherjee
authors.affiliation: International Centre for Genetic Engineering and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India pawanmal@gmail.com sunilm@icgeb.res.in raj@icgeb.res.in.
authors.forename: Raj K
authors.initials: RK
authors.lastname: Bhatnagar
mesh_terms: D000217:Acyltransferases; D002268:Carboxypeptidases / Q000235:genetics / Q000378:metabolism; D003715:Dengue / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D003716:Dengue Virus / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D050883:HSC70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D056918:Nuclear Receptor Coactivators / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D062805:Synaptogyrins / Q000096:biosynthesis / Q000235:genetics; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics; D000085242:Viral Proteases
chemical_list: D000217:Acyltransferases; D002268:Carboxypeptidases; D050883:HSC70 Heat-Shock Proteins; D035683:MicroRNAs; D056918:Nuclear Receptor Coactivators; D012697:Serine Endopeptidases; D062805:Synaptogyrins; D014157:Transcription Factors; C494371:HSPA8 protein, human; C486559:NCOA6 protein, human; C507923:SYNGR1 protein, human; C108675:TAFAZZIN protein, human; C436327:SCPEP1 protein, human; C504449:NS3 protease, dengue virus; D000085242:Viral Proteases
keywords: HSC70; dengue virus; miRNAs; viral suppressors of RNA silencing (VSR)