Searching for cellular factors interacting with influenza A virus PA-X protein

• Main Authors: Cheng-Huei Hung, 洪呈輝
• Other Authors: Shih-Yen Lo
• Format: Others
• Language: zh-TW
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/76703841587589075166

碩士 === 慈濟大學 === 醫學生物技術碩士班 === 103 === Influenza A viruses are negative-sense, single-stranded, segmented RNA viruses. The virus particle is 80–120 nanometers in diameter and usually roughly spherical, although some rare filamentous occurs. Influenza A virus (IAV) infection leads to variable and imperfectly understood pathogenicity. It’s reported that segment 3 of the virus contains a second open reading frame (“X-ORF”), generated via ribosomal frameshifting. The frameshift product, termed PA-X, is encoded by the X-ORF and has functions to repress cellular gene expression. PA-X also modulates IAV virulence in a mouse infection model, acting to decrease pathogenicity. Loss of PA-X expression leads to changes in the kinetics of the global host response. Because of its influence with host immunopathoogy, PA-X may be a fruitful target for new therapeutics of influenza A virus. Here we use yeast-two-hybrid system to investigate the factors in human cell that interact with PA-X. With HL4024 cDNA library, which is reverse transcribed from liver cell and cloned into pACT2 vector, we have found an important protein RBP4. Retinol binding protein 4(RBP4) belongs to the lipocalin family and is the specific carrier for retinol (vitamin A alcohol) in the blood. It delivers retinol from the liver stores to the peripheral tissues. RBP4 is recently studied as the marker of hepatitis c virus cirrhosis and steatosis. Confocal microscopy and co-immunoprecipitation assays are needed to prove that RBP4 does interact with PA-X in A549 cell.