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Objectives: Identify from among proteomics data host factors that modulate influenza viral polymerase activity by functional genomics, map protein-protein interactions and mechanisms of modulation on the viral life cycle (validation of proteomics and functional genomics), and strain-specific differences. Adapt exBiFC assay for co-localization studies (PMID: 17191610). Contribute to DFIM, structural studies on M1 and NP (in collaboration), and vPOL epidemiological and functional modeling.
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Interaction of viral polymerase/NP with 2 cellular proteins: P1 and F1

Trial 1 Westerns (0607):

Results: Fig. 1. P1 binds HA7-agarose (not precleared); F1 shows no interaction in IP from transfectants. Repeat experiment with new transfectants, preclearing, varying buffers.

Trial 2 Westerns (0707):

Results: Interactions of P1, F1, and H1 with NP. All transfectant lysates precleared before immunoprecipitation.
Fig. 2. P1 interacts with vPOL by NP-FLAG immunoprecipitation, but weakly with NP-HA alone; low levels of P1 in NP-HA +3P transfection preclude further conclusion as yet (left). Binding of F1 to NP is stabilized by viral pol subunits PB1,PB2,PA (right); or possibly NP-FLAG.

Fig. 3. Protein H1 binds to NP.

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