Data from: Linking germline telomere removal to global programmed DNA elimination in Tetrahymena genome differentiation
Data files
Jul 09, 2026 version files 398.96 MB
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Law_data_Fig2-Sup1.zip
97.46 MB
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Law_Data_Fig2.zip
70.97 MB
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Law_data_Fig4_Sup1.zip
46.92 MB
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Law_Data_Fig4.zip
79.18 MB
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Law_Data_Fig5.zip
30.66 MB
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Law_Data_Fig6.zip
39.05 MB
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Law_data_Fig8.zip
34.72 MB
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README.md
2.95 KB
Abstract
In the ciliate Tetrahymena, telomeres of the germline micronucleus (MIC) are removed and replaced by de novo telomere addition during somatic macronuclear (MAC) development. In this study, we investigated the kinetics and mechanism of the MIC telomere elimination. Comparison of the MIC and MAC genome sequences indicated that the MIC telomeres are excised from chromosomes as part of larger MIC-limited sequences (MLSs) through chromosomal breakage. We confirmed this using an optimized oligo-FISH protocol and and found that their elimination occurs in parallel with other programmed DNA elimination processes. CRISPR-Cas9 disruption of a MLS-associated Chromosome Breakage Sequence (CBS) showed that elimination of the MLS was not blocked but instead led to loss of adjacent MAC-destined sequence (MDS), suggesting abnormal co-elimination. In biparental crosses of the CBS mutant, however, both MLS and MDS were retained, DNA elimination was broadly disrupted, and no viable progeny were produced. These findings indicate that chromosome breakage at MLS-associated CBSs is essential for the proper separation of MLSs and MDSs, ensuring correct DNA elimination and successful sexual progeny development. We propose that the MIC telomere elimination is subsumed within the broader process of programmed DNA elimination.
Dataset DOI: 10.5061/dryad.2z34tmq1w
Description of the data and file structure
Files and variables
File: Law_data_Fig8.zip
Description: Exconjugants at 32 hours post-mixing (hpm) or conjugating pairs at 9 hpm from a cross between two wild-type strains (WT x WT) and two independent crosses between a wild-type and a 4R-CBS mutant strain (WT x Mut, Cross 1 and Cross 2) were analyzed. FISH was performed using probes complementary to the MIC-specific Tlr1 element, and immunofluorescence staining was performed using an antibody against di- and tri-methylated histone H3 at lysine 9 (H3K9me, green). DNA was counterstained with DAPI.
File: Law_Data_Fig6.zip
Description: Exconjugants at 30 hours post-mixing (hpm) from crosses between two wild-type strains (WT x WT), two 4R-CBS mutant strains (Mut x Mut), or a wild-type and a 4R-CBS mutant strain (WT x Mut) were analyzed by oligo-FISH using a pool of oligonucleotide probes complementary to 4R-MLS and 4R-MDS . DNA was counterstained with DAPI.
File: Law_data_Fig4_Sup1.zip
Description: Conjugating wild-type cells at the indicated time points (hours post-mixing, hpm) were analyzed by oligo-FISH using pool of oligonucleotide probes complementary to 3L-MLS. DNA was counterstained with DAPI.
File: Law_Data_Fig2.zip
Description: Vegetative (Veg) and conjugating wild-type cells at the indicated time points (hours post-mixing, hpm) were analyzed by oligo-FISH using an oligonucleotide probe complementary to the MIC-specific G4T3 repeat. DNA was counterstained with DAPI.
File: Law_Data_Fig4.zip
Description: Vegetative (Veg) and conjugating wild-type cells at the indicated time points (hours post-mixing, hpm) were analyzed by oligo-FISH using pool of oligonucleotide probes complementary to 4R-MLS or 4R-MDS. DNA was counterstained with DAPI.
File: Law_Data_Fig5.zip
Description: Genomic DNA was extracted from wild-type cells under starvation and during conjugation were used for the telomere-anchored PCR assay. The coding region of RPL11 was also amplified from the same genomic DNA by single-step PCR as a control. Telomere-capped 4R-MLS and 3L-MLS ends are expected to produce PCR products of approximately 269 bp and 125 bp, respectively (Fig. 5D). Two independent crosses of 4R-CBS mutants during conjugation were also analyzed (Fig. 5E).
File: Law_data_Fig2-Sup1.zip
Description: Conjugating wild-type cells at the indicated time points (hours post-mixing, hpm) were analyzed by FISH using probes complementary to the MIC-specific Tlr1 element. DNA was counterstained with DAPI.
Code/software
The ".czi" files can be opened and viewed by Fiji (https://imagej.net/software/fiji/).
