Cell fusion

UC Berkeley–UC San Francisteo Graduate Group in Bioengineering, United States; Department of Bioengineering & Biophysics Graduate Group, University of California, Berkeley, United States; Division of Biological Systems and Engineering, Lawrence Berkeley National Laboratory, United States; Chan Zuckerberg Biohub, United States;


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Cell-cell fusion, which is essential for tissue development và used by some viruses khổng lồ form pathological syncytia, is typically driven by fusogenic membrane proteins with tall (>10 nm) ectodomains that undergo conformational changes khổng lồ bring apposing membranes in cthua thảm tương tác prior to lớn fusion. Here we report that a viral fusoren with a short (

Introduction





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Expression of p14 drives cell-cell fusion & is quantified with a splitYFP fluorescent assay.


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(a) Schematic of fusion-associated small transmembrane protein, p14, in proportion to post-fusion trimeric structure of cell-cell fusogens, Eff-1 (PDB:4OJC) và Hap2 (PDB: 5MF1), on the plasma membrane. (b) Expression of p14 in HEK293T cells drives cell-cell fusion forming large multinucleated cells that increases kích cỡ và number of nuclei over time. (c) Average nuclei count of HEK293T cells expressing p14 at 12 hr & 24 hr. P-values are ks-test and error bars represent standard deviation from 3 independent transfections (See also Figure 1—source data 1). (d) Percent of p14 expressing nuclei in cells with 3 or more nuclei at 12 hr and 24 hr. P-values are two-tailed, two-sample Student’s t-thử nghiệm where ** = pYFP of sample is the YFP fluorescence intensity of the sample, IYFP.. of sample is the YFP fluorescence intensity of the non-transfected cells, and IYFPhường of sample is the YFP fluorescence intensity of cells transfected with p14 WT. (f) Average YFPhường fluorescence intensity of HEK293T cells expressing p14 at 12 hr và 24 hr with error bars representing standard deviations from 3 independent transfections of 3 wells each. P-values are two-tailed, two-sample Student’s t-test where *** = pFigure 1—figure supplement 1c, (d). (g) Average fusion index of p14 cytoplasmic truncation mutant with error bars representing standard deviations from 3 independent transfections of 3 wells each. P-values are two-tailed, two-sample Student’s t-kiểm tra where *** = pFigure 1—figure supplement 1e, (f).



Download elife-51358-fig1-data1-v2.xlsx

Results




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