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Hepatitis B Virus X Protein Regulate Sprouty Homolog 1 Activation of MAPK Signaling Pathway

Author: LiuWei
Tutor: LiaoShiQi
School: Lanzhou University of Technology
Course: Biochemistry and Molecular Biology
Keywords: Hepatitis B virus HBx MAPK signaling pathway SPRY1 Apoptosis Cell proliferation HEK293 cells
CLC: R373
Type: Master's thesis
Year: 2010
Downloads: 75
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Abstract


In the genome of the hepatitis B virus (HBV) X gene and its encoded product X protein (hepatitis B virus X protein, HBx) plays an important role in the regulation of HBV replication. HBx involved in a number of cellular signal transduction pathways, cell proliferation, apoptosis play an important regulatory role. It is mainly in conjunction with some key cell signaling pathways regulating protein complete the regulatory role of cellular signal transduction pathway. These regulatory role, including activation of host cells and viral gene transcription, regulation of apoptosis, inhibition of the cells damaged DNA exonuclease repair the reaction and activate cellular signal transduction pathway if mitosis the original (MAPK), JAK, STAT-level linking reaction and the like. HBx involved in many signal transduction pathways, we are concerned about HBx activation of MAPK signal transduction pathway, this pathway is a highly conserved module-related cell functions, including cell proliferation, differentiation, and migration, and tumor formation and play a huge role in regulating the migration process. But not currently found in the direct role of MAPK signaling in HBx protease receptor to activate the potential molecular mechanisms remain elusive. Through the research of this subject to preliminary clarify HBx encoded proteins in the liver of MAPK inhibition receptor --- SPRY1 gene encoding the protein to activate MAPK signal transduction pathway by regulating. In order to clarify this mechanism, the subject has taken a series of experimental studies: (1) Construction of Flag-HBx, Myc-SPRY1 Myc-SPRY1-53-Mut eukaryotic expression vector; (2) determine Flag-HBx and Myc-SPRY1,, protein of between the existence of interactions; (3) to explore the Flag-HBx protein encoded with the Myc-SPRY1 encoding protein interaction domain; (4) the Myc-SPRY1 encoding proteins whether the impact of the Flag-HBx protein in mammalian HEK293 cells expression levels; (5) of the AP-1 luciferase reporter gene Experimental Study of Flag-HBx the Myc-SPRY1 encoding proteins of the AP-1 transcription factor; (6) research Flag-HBx and the Myc-SPRY1 encoding proteins on HEK293 cell apoptosis and proliferation of functional impact. The conclusions are as follows: 1. Successfully constructed Flag-HBx Myc-SPRY1, the Myc-SPRY1 of-53-Mut, the eukaryotic expression vector; 2 by co-immunoprecipitation (Co-IP) and Pull-down experiments to determine the Flag-HBx and the Myc-SPRY1 exist in mammalian HEK293 cells in vitro and in vivo interaction; 3 by co-immunoprecipitation experiments showed that the tyrosine at SPRY1 53 mutant does not affect the interaction between them, but the AP-1 luciferase reporter genetic experiments show that the of SPRY1 53 tyrosine mutants affect cellular signal transduction pathways with HBx activation, so we speculate that their interaction region including a sequence; 4. Myc-SPRY1 encode proteins affecting Flag-HBx encoding the expression level of the protein in the mammalian HEK293 cells, and the levels of the protein encoded by the Flag-HBx With the increase in protein expression levels of the Myc-SPRY1 coding increases; 5. Flag-HBx synergistic Myc-SPRY1 nuclear transcription factor AP-1-luc transcriptional activity of having an active effect; 6 through apoptosis assay and cell proliferation experiments and found that Flag-HBx promote proliferation of HEK293 cells, inhibition of HEK293 cells apoptosis; and found that when the co-transfection of Myc- into HEK293 cells, SPRY1 and Flag-HBx, Myc-SPRY1 increased Flag-HBx anti-apoptotic effects of HEK293 cells and the Myc-SPRY1 raised the Flag-HBx role HEK293 cells proliferation level.

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CLC: > Medicine, health > Basic Medical > Medical Microbiology ( pathogenic bacteriology,pathogenic microbiology ) > Human Virology ( pathogenic virus)
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