title: Role of human GRP75 in miRNA mediated regulation of dengue virus replication.
abstract: In recent times, RNAi has emerged as an important defence system that regulates replication of pathogens in host cells. Many RNAi related host factors especially the host miRNAs play important roles in all intrinsic cellular functions, including viral infection. We have been working on identification of mammalian host factors involved in Dengue virus infection. In the present study, we identified Glucose Regulated Protein 75kDa (GRP75), as a host factor that is associated with dicer complex, in particular with HADHA (trifunctional enzyme subunit alpha, mitochondrial), an auxiliary component of dicer complex. Knockdown of GRP75 by respective siRNAs in Huh-7 cells resulted in the accumulation of dengue viral genomic RNA suggesting a role of GRP75 in regulating dengue virus replication in human cell lines. To elucidate the mode of action of GRP75, we over expressed the protein in Huh-7 cells and analysed the host miRNAs processing. The results revealed that, GRP75 is involved in processing of host miRNA, hsa-mir-126, that down regulates dengue virus replication. These findings suggest a regulatory role of human miRNA pathway especially GRP75 protein and hsa-mir-126 in dengue virus replication. These results thus provide insights into the role of miRNAs and RNAi machinery in dengue life cycle.
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: Translational Health Science and Technology Institute, NCR Biotech Science Cluster, 3rd Milestone, Faridabad-Gurgaon Highway, PO Box # 04, Faridabad 121001, India.
authors.forename: Guruprasad R
authors.initials: GR
authors.lastname: Medigeshi
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. Electronic address: pawanmal@gmail.com.
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. Electronic address: sunilm@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. Electronic address: raj@icgeb.res.in.
authors.forename: Raj K
authors.initials: RK
authors.lastname: Bhatnagar
mesh_terms: D045744:Cell Line, Tumor; D003716:Dengue Virus / Q000502:physiology*; D057809:HEK293 Cells; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism*; D012323:RNA Processing, Post-Transcriptional; D043244:Ribonuclease III / Q000378:metabolism; D014779:Virus Replication
chemical_list: D018840:HSP70 Heat-Shock Proteins; C492001:HSPA9 protein, human; C527111:MIRN126 microRNA, human; D035683:MicroRNAs; D024101:Mitochondrial Proteins; D043244:Ribonuclease III
keywords: Dengue virus; Dicer; GRP75; HADHA; miRNA