The mitochondrial carrier homolog 2 is involved in down-regulation of influenza A virus replication

• Published in: Molecular biology reports Vol. 51; no. 1; p. 642
• Main Authors: Ulupinar, Pınar, Çağlayan, Elif, Rayaman, Erkan, Nagata, Kyosuke, Turan, Kadir
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.12.2024; Springer Nature B.V
• Subjects: Amino acids; Animal Anatomy; Animal Biochemistry; Apoptosis; Biomedical and Life Sciences; Down-Regulation; Glutathione; HEK293 Cells; HeLa Cells; Histology; Homeostasis; Humans; Influenza; Influenza A; Influenza A virus - genetics; Influenza A virus - physiology; Life Sciences; Lipid metabolism; Membrane proteins; Mitochondria; Mitochondria - metabolism; Mitochondrial Membrane Transport Proteins - genetics; Mitochondrial Membrane Transport Proteins - metabolism; Mitochondrial Proteins - genetics; Mitochondrial Proteins - metabolism; Morphology; Original Article; Protein Binding; Protein turnover; Proteins; Replication; RNA-Dependent RNA Polymerase - genetics; RNA-Dependent RNA Polymerase - metabolism; siRNA; Viral Proteins - genetics; Viral Proteins - metabolism; Virus Replication - genetics; Viruses; Influenza PA protein; MTCH2; Viral RdRp; Yeast two hybrid; Protein–protein docking
• ISSN: 0301-4851; 1573-4978
• DOI: 10.1007/s11033-024-09584-5

Background The mitochondrial carrier homolog 2 (MTCH2) is a mitochondrial outer membrane protein regulating mitochondrial metabolism and functions in lipid homeostasis and apoptosis. Experimental data on the interaction of MTCH2 with viral proteins in virus-infected cells are very limited. Here, the interaction of MTCH2 with PA subunit of influenza A virus RdRp and its effects on viral replication was investigated. Methods The human MTCH2 protein was identified as the influenza A virus PA-related cellular factor with the Y2H assay. The interaction between GST.MTCH2 and PA protein co-expressed in transfected HEK293 cells was evaluated by GST-pull down. The effect of MTCH2 on virus replication was determined by quantification of viral transcript and/or viral proteins in the cells transfected with MTCH2-encoding plasmid or MTCH2-siRNA. An interaction model of MTCH2 and PA was predicted with protein modeling/docking algorithms. Results It was observed that PA and GST.MTCH2 proteins expressed in HEK293 cells were co-precipitated by glutathione-agarose beads. The influenza A virus replication was stimulated in HeLa cells whose MTCH2 expression was suppressed with specific siRNA, whereas the increase of MTCH2 in transiently transfected HEK293 cells inhibited viral RdRp activity. The results of a Y2H assay and protein-protein docking analysis suggested that the amino terminal part of the viral PA (nPA) can bind to the cytoplasmic domain comprising amino acid residues 253 to 282 of the MTCH2. Conclusion It is suggested that the host mitochondrial MTCH2 protein is probably involved in the interaction with the viral polymerase protein PA to cause negative regulatory effect on influenza A virus replication in infected cells.