Fig. 6: Increased DNA damage promotes RAD51 interaction with host HOP2 but not viral Rep. | Nature Communications

Fig. 6: Increased DNA damage promotes RAD51 interaction with host HOP2 but not viral Rep.

From: H3.1K27me1 loss confers Arabidopsis resistance to Geminivirus by sequestering DNA repair proteins onto host genome

Fig. 6

a BiFC of YFP assays showed that mutations of ATXR5/6 promote the interaction between RAD51 and HOP2. See Supplementary Fig. 13d for negative controls. 15 independent protoplasts were examined for the interaction and showed similar results. Scale bars, 5 μm. b Statistical analysis of YFP signal intensity of each protoplast. c BiFC of YFP assays showed that interaction between RAD51 and Rep was compromised in atxr5 atxr6. See Supplementary Fig. 13e for negative control. 15 independent protoplasts were examined for the interaction and showed similar results. Scale bars, 5 μm. d Statistical analysis of YFP signal intensity of each protoplast. In (b) and (d), each dot in the bar plot represents one protoplast. Data are presented as mean ± SD (n = 15 biologically independent prototplasts. e q-PCR assays show the amount of rDNA in different lines. The relative amount of rDNA was normalized against UBQ10. f Representative phenotypes of CaLCuV-inoculated Col-0, atxr5 atxr6, and third generation of (T3) lines expressing H3.1 variants. Photographs were taken at 14 dpi. Scale bars, 1 cm. g Percentages of symptomatic plants induced by CaLCuV-inoculation in indicated backgrounds at 10, 12, and 14 dpi. h q-PCR assays show the amount of viral DNA A in CaLCuV-inoculated plants at 14 dpi. Normalization of viral DNA was conducted as Fig. 2g. i SA treatment activates the DNA damage response. qRT-PCR assays show the transcription level of HRR factors in indicated genotypes and treatments. j q-PCR assays show the relative amount of viral DNA A in the indicated genotypes and treatments at 14 dpi. In (e), (g), (h), (i), (j), each dot in the bar plot represents one replicate. Experiments were performed with 36 plants/replicate except in (i) 6 plants/replicate was used. Data are presented as mean ± SD (n = 3 biological replicates). Statistics In Figs. 6b, d, e, g, h, i and j were performed with unpaired two-tailed student t-test, *, **, *** and ****, P < 0.05, 0.01, 0.001, and 0.0001, respectively. Source data are provided in Source Data File.

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