Data for: The CD4 transmembrane GGXXG and juxtamembrane (C/F)CV+C motifs mediate pMHCII-specific signaling independently of CD4-Lck interactions
Data files
Mar 15, 2024 version files 302.06 KB
Abstract
We previously reported that the transmembrane GGXXG motif co-arose with the cytoplasmic juxtamembrane (C/F)CV+C motif in eutherian (placental mammal) CD4 and that constituent residues therein evolve under purifying selection (Lee, et al., 2022). These data led us to discover that mutating these motifs together increased CD4-Lck association but reduced CD3z, Zap70, and Plcg1 phosphorylation levels as well as IL-2 production. Because these mutants preferentially localized CD4-Lck pairs to non-membrane raft fractions, one potential explanation for our results was that they impaired proximal signaling by sequestering Lck away from TCR-CD3. An alternative hypothesis is that the mutations directly impacted TCR-CD3 signaling because the wild type motifs play a Lck-independent role in signaling. Here we used a reductionist approach to discriminate between these possibilities (Lee, et al., 2023). Our current data lead us to conclude that: intracellular CD4-Lck interactions are not necessary for pMHCII-specific signal initiation; the GGXXG and (C/F)CV+C motifs are key determinants of CD4-mediated pMHCII-specific signal amplification; the GGXXG and (C/F)CV+C motifs exert their functions independently of direct CD4-Lck association. These data provide a mechanistic explanation for why residues within these motifs are under purifying selection. The results are also important to consider for biomimetic engineering of synthetic receptors.
README: Data for The CD4 transmembrane GGXXG and juxtamembrane (C/F)CV+C motifs mediate pMHCII-specific signaling independently of CD4-Lck interactions
Data associated with the statistical analyses described. Each figure has a separate file.
Data was exported from Prism
Description of the data and file structure
Data associated with the statistical analyses described in the paper. Each figure has a separate file. specific figure is indicated in the title of the file
Data was exported from Prism and is provides as an excel file.
Figure_2_stats_export.xlsx: contains the raw values used for the experiment described in Figure 2
SFig_3_stat_export.xlsx: contains the raw values used for the experiment described in Supplementary Figure 3
Figure_3_stats_export.xlsx: contains the raw values used for the experiment described in Figure 3
Figure_4_stats_export.xlsx: contains the raw values used for the experiment described in Figure 4
Figure_5_stats_export.xlsx: contains the raw values used for the experiment described in Figure 5
SFig_2_Stats_export.xlsx: contains the raw values used for the experiment described in Supplementary Figure 2
SFig_5_stats_export.xlsx: contains the raw values used for the experiment described in Supplementary Figure 5
SFig_4_stat_export.xlsx: contains the raw values used for the experiment described in Supplementary Figure 4
master_intensity_and___responders_work_sheet_for_normalization.xlsx: Describes the logic used to normalize the data
Methods
Methods are described in the manuscript
Usage notes
excel or any text editor