Molecular mechanism of the hepatitis C vitus core protein chaperone properties : physicochemical investigation by fluorescence and surface plasmon resonance
SHARMA, Kamal Kant
Molecular mechanism of the hepatitis C vitus core protein chaperone properties : physicochemical investigation by fluorescence and surface plasmon resonance.
Thèses de doctorat,
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Open reading frame (ORF) of 9.6kb HCV genomic RNA encodes at least 10 proteins, 4 structural and 6 non-structural, during translation process. The core is one of those 4 structural proteins and considered as a multifunctional chaperone involving in several viral processes like cell proliferation, differentiation, RNA packaging, nucleocapsid formation and recombinant genetic variability. With the virtue of its chaperone properties, Domain D1 dimerises the 3’ untranslated region (3’ UTR) of the genomic RNA. However, the mechanism of the core chaperone activity in the dimerisation of the genomic RNA and in binding with its target nucleic acids are still unknown and were investigated in this present project. To reach this objective, we used fluorescence and surface plasmon resonance (SPR) techniques. By using the native D1 domain and peptides derived from this domain, we first characterized the binding parameters and the conformational changes associated with the binding of these peptides to the native and mutated sequences from HCV 3’ UTR sequences. Next, we investigated the destabilization of model and HCV ODNs secondary structure by the D1 domain and its mutants and found that core peptides only marginally destabilise the secondary structures of ODNs. In a last step, we described the molecular mechanisms of the core chaperone properties based on the hybridization kinetics of various HCV and model oligonucleotides. These chaperone properties of core are thought to intervene in processes like the encapsidation, the synthesis of the complementary strand of the genomic RNA and the recombination mechanisms participating to the genetic variability of the virus.
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