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. 2024 Sep 3;15:7653. doi: 10.1038/s41467-024-52147-2

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

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Fig. 4 — a Analysis of TRP1-mtDNA transfer to nucleus. Wild-type, nuc1Δ and nuc1-H138A mutant cells replica plated from rich YEPD media onto plates with media lacking tryptophan. Trp⁺ colonies represent transfer of TRP1-mtDNA to nucleus. b Frequency of Trp⁺ colonies in wild-type, nuc1Δ, and nuc1-H138A mutant cells, (mean ± SD; n = 7 for 0 or 3 days and 11 for 8 days for wild type. n = 3 for 0 or 3 days, and 6 for 8 days for nuc1Δ mutant. n = 4 for 0 or 3 days, and 7 for 8 days for nuc1-H138A mutant. n represents biological repeats. P values determined using unpaired two-tailed Welch’s t-test). c Model of Nuc1 mediated control of mtDNA transfer to the nucleus. In stationary phase cells, mtDNA is released from mitochondria in Nuc1-independent manner. MtDNA is degraded by Nuc1 nuclease. Incomplete degradation leaves small mtDNA pieces that can be integrated at DSBs in nuclear genome. In the absence of Nuc1, mtDNA is not degraded and large mtDNA pieces are transferred to the nucleus where they either form circular mtDNA of different sizes or rarely integrate in the genome. d Southern blot analysis and quantification of long mtDNA and mtDNA smear in wild-type and nuc1Δ at 8 days stationary phase cells, (mean ± SD; n = 4; n represents biological repeats. P values determined using unpaired two-tailed Welch’s t-test). e Random spore analysis on plates with media lacking tryptophan. Trp⁺ colonies represent transfer of TRP1-mtDNA to nucleus. Frequency of spores carrying nuclear TRP1-mtDNA is shown. (mean ± SD; n = 3 for wild type and 4 for nuc1Δ mutant; n represents biological repeats. P values determined using unpaired two-tailed Welch’s t-test). f Scheme and analysis of insertion originating from transformed DNA fragments of different sizes. Frequency of insertions of annealed oligonucleotides (24–74 bp) or PCR fragments (84–1000 bp) are shown. 480–1680 independent NHEJ products were tested by PCR. Source data are provided as a Source Data file.