Replication timing of large Sorex granarius (Soricidae, Eulipotyphla) telomeres
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Previously, we described the unique feature of telomeric regions in Iberian shrew Sorex granarius: its telomeres have two ranges of size, very small (3.8 kb of telomeric repeats on average) and very large discontinuous telomeres (213 kb) interrupted with 18S rDNA. In this study, we have demonstrated extraordinary replication pattern of S. granarius large telomeres that have not been shown before in other studied mammal. Using the ReD-FISH procedure, we observed prolonged, through S period, large telomere replication. Furthermore, revealed ReD-FISH asymmetric signals were probably caused by partial replication of telomeres within an hour of 5-bromodeoxyuridine treatment due to the large size and special organization. We also found that in contrast to the telomeric halo from primary fibroblasts of bovine, mink, and common shrew, telomere halo of S. granarius consists of multiple loops bundled together, some of which contain rDNA. Here, we suggested several replicons firing possibly stochastic in each large telomere. Finally, we performed the TIF assay to reveal DNA damage responses at the telomeres, and along with TIF in nuclei, we found large bodies of telomeric DNA and ɤ-H2AX in the cytoplasm and on the surface of fibroblasts. We discuss the possibility of additional origin activation together with recombination-dependent replication pathways, mainly homologous recombination including BIR for replication fork stagnation overcoming and further S. granarius large telomere replication.
KeywordsTelomere replication ReD-FISH Halo Sorex granarius
Fluorescence in situ hybridization
Replication detargeting FISH
- PNA probe
Peptide nucleic acid probe
Fluorescence-activated cell sorting
Telomere dysfunction-induced foci
This work was supported by budget project no. 0324-2018-0019 of the Federal Research Center Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences.
The experiments comply with the current laws of Russian Federation country in which they were performed.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Studies with human or animal
All animal studies were undertaken with prior approval from Interinstitutional Bioethical Committee of ICG SB RAS.
This article does not contain any studies with human or animal subjects performed by the any of the authors.
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