Fig. 7: A polybasic domain outside the SNARE motif is critical for tomosyns’ function. | Nature Communications

Fig. 7: A polybasic domain outside the SNARE motif is critical for tomosyns’ function.

From: Tomosyns attenuate SNARE assembly and synaptic depression by binding to VAMP2-containing template complexes

Fig. 7

a Schematic representations of wild-type and mutant tomosyn-1m. The amino acid (aa) numbers and mutation site are stated on the right. b Example traces of 10 Hz. c STP quantified by the ratio of the fifth pulse over the first pulse. Control n = 21/6, cDKO n = 25/8, +WT n = 27/6, +FA n = 18/5, +ΔPB n = 13/4, +ΔSNARE n = 8/4, +WDonly n = 10/3. Post hoc comparisons against cDKO: ***p = 0.0001 (control); ***p < 0.0001 (+WT), p = 0.3013 (+FA), p = 0.8781 (+ΔPB), p = 0.7821 (+ΔSNARE), p = 0.2592 (+WDonly). d–e A recovery pulse (R) was given 2 s after 40 Hz stimulation. d Example traces. e Quantification of EPSC recovery (normalized to 1st). Control n = 21/6, cDKO n = 25/8, +WT n = 25/6, +FA n = 16/5, +ΔPB n = 12/4, +ΔSNARE n = 6/3, +WDonly n = 9/2. Post hoc comparisons against cDKO: ***p = 0.0001 (control); ***p < 0.0001 (+WT), p = 0.03677 (+FA), p = 0.4757 (+ΔPB), p = 0.6820 (+ΔSNARE), p = 0.8506 (+WDonly). f, g Immunostainings confirm expression of all mutants. f Example images of dendrites. Scale bar = 10 µm. g Synaptic tomosyn-1 expression normalized to control within each neuronal culture. Control n = 26/3, cDKO n = 27/3, +WT n = 29/3, +FA n = 33/3, +ΔPB n = 30/3, +ΔSNARE n = 36/3, +WDonly n = 34/3. Post hoc comparisons against +WT: p = 0.1731 (control); ***p < 0.0001 (cDKO), ***p < 0.0001 (+FA), p = 0.0461 (+ΔPB), ***p < 0.0001 (+ΔSNARE), p = 0.0005 (+WDonly). Post hoc comparisons against cDKO: ***p < 0.0001 (control); ***p < 0.0001 (+WT), ***p < 0.0001 (+FA), ***p < 0.0001 (+ΔPB), ***p < 0.0001 (+ΔSNARE), ***p < 0.0001 (+WDonly). N = cells/independent cultures. In (c, e, g), boxplots display median (center), upper and lower quartiles (box bounds) and whiskers to the last datapoint within 1.5x interquartile range. A one-way ANOVA tested the significance of adding experimental group as a predictor, see Supplementary Table 1. For post-hoc comparisons, p-value thresholds (*<0.05; **<0.01;***<0.001) were adjusted with a Bonferroni correction. Red asterisks show comparison to cDKO, gray asterisks show comparison to + WT. Abbreviations: PB (polybasic domain), n.s. (not significant). See also Supplementary Fig. 11. Source data are provided as a Source Data file.

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