tailieunhanh - Báo cáo y học: "Biophysical analysis of HTLV-1 particles reveals novel insights into particle morphology and Gag stoichiometry"

Tuyển tập các báo cáo nghiên cứu về y học được đăng trên tạp chí y học 'Respiratory Research cung cấp cho các bạn kiến thức về ngành y đề tài: Biophysical analysis of HTLV-1 particles reveals novel insights into particle morphology and Gag stoichiometry. | Grigsby et al. Retrovirology 2010 7 75 http content 7 1 75 RETR0VIR0L0GY RESEARCH Open Access Biophysical analysis of HTLV-1 particles reveals novel insights into particle morphology and Gag stoichiometry Iwen F Grigsby1 2 Wei Zhang1 2 Jolene L Johnson1 4 Keir H Fogarty1 4 Yan Chen1 4 Jonathan M Rawson1 Aaron J Crosby1 Joachim D Mueller1 4 Louis M Mansky1 2 3 Abstract Background Human T-lymphotropic virus type 1 HTLV-1 is an important human retrovirus that is a cause of adult T-cell leukemia lymphoma. While an important human pathogen the details regarding virus replication cycle including the nature of HTLV-1 particles remain largely unknown due to the difficulties in propagating the virus in tissue culture. In this study we created a codon-optimized HTLV-1 Gag fused to an EYFP reporter as a model system to quantitatively analyze HTLV-1 particles released from producer cells. Results The codon-optimized Gag led to a dramatic and highly robust level of Gag expression as well as virus-like particle VLP production. The robust level of particle production overcomes previous technical difficulties with authentic particles and allowed for detailed analysis of particle architecture using two novel methodologies. We quantitatively measured the diameter and morphology of HTLV-1 VLPs in their native hydrated state using cryotransmission electron microscopy cryo-TEM . Furthermore we were able to determine HTLV-1 Gag stoichiometry as well as particle size with the novel biophysical technique of fluorescence fluctuation spectroscopy FFS . The average HTLV-1 particle diameter determined by cryo-TEM and FFS was 71 20 nm and 75 4 nm respectively. These values are significantly smaller than previous estimates made of HTLV-1 particles by negative staining TEM. Furthermore cryo-TEM reveals that the majority of HTLV-1 VLPs lacks an ordered structure of the Gag lattice suggesting that the HTLV-1 Gag shell is very likely to be organized differently compared to .

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