Rif1 regulates telomere length through conserved HEAT repeats

Shubin, Calla B; Mayangsari, Rini; Swett, Ariel D; Greider, Carol W.

doi:10.1093/nar/gkab206

Cited by 9 publications

(18 citation statements)

References 44 publications

(39 reference statements)

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“…This small effect is of interest, as it suggests a potential additional role of Rif1 protein on telomere length. Rif1 can exert telomere specific length regulation when it is tethered at a unique telomere (Shubin et al 2021).…”

Section: Telomere Shortening In the Absence Of Telomerase Reveals The Minimal Functional Telomerementioning

confidence: 99%

“…Yeast molecular cloning, culturing, sporulation, and transformation were performed as previously described in (Green and Sambrook 2012;Keener et al 2019;Shubin et al 2021).…”

Section: Molecular Cloningmentioning

confidence: 99%

“…The UE1 (previous named 1L 5XUAS landing pad) and UE11 (previously named 11R 5XUAS landing pad) constructs were designed in silico using SnapGene software and constructed as described in (Shubin et al 2021). UE11 was generated in the same manner as UE1, but with homology at chromosome arm 11R, rather than chromosome arm 1L.…”

Section: Molecular Cloningmentioning

confidence: 99%

“…CVy61 (W303-0 WT haploids), UE1 (previously CALy117, 1L::leu2), UE11 (previously CALy119, 11R::leu2). UE1 rif1 (previously CALy202, 1L::leu2, rif1::kanMX), UE11 rif1 (previously CALy204, 11R::leu2, rif1::kanMX), and UE1 tel1 (previously CALy472, 1L::leu2, tel1:: hphMX4) strains were generated as previously described in (Shubin et al 2021). These strains were re-named UE1 and UE11 to highlight the function of a unique end, rather than a 5XUAS landing pad, that was important to this study.…”

Section: Yeast Strains Culturing and Transformationmentioning

confidence: 99%

“…After transfer, the membrane was hybridized with 32 P radiolabeled 2-log ladder and PCR fragments unique to either 1L, 11R (a purified PCR product of the LEU2 gene and CYC1 terminator as described in Shubin, et al), Y' subtelomeric elements as described in Kaizer, et al or to CEN4 (Kaizer et al 2015;Laterreur et al 2018;Shubin et al 2021). After washing, the membrane was exposed on a Storage Phosphor screen (GE Healthcare) for 1 day for bulk assays and 4-5 days for single telomere assays.…”

Section: Southern Blotting and Densitometrymentioning

confidence: 99%

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Chromosome specific telomere lengths and the minimal functional telomere revealed by nanopore sequencing

Sholes

Karimian

Gershman

et al. 2021

Preprint

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We developed a method to tag telomeres and measure telomere length by nanopore sequencing in the yeast S. cerevisiae. Nanopore allows long read sequencing through the telomere, subtelomere and into unique chromosomal sequence, enabling assignment of telomere length to a specific chromosome end. We observed chromosome end specific telomere lengths that were stable over 120 cell divisions. These stable chromosome specific telomere lengths may be explained by stochastic clonal variation or may represent a new biological mechanism that maintains equilibrium unique to each chromosomes end. We examined the role of RIF1 and TEL1 in telomere length regulation and found that TEL1 is epistatic to RIF1 at most telomeres, consistent with the literature. However, at telomeres that lack subtelomeric Y' sequences, tel1Δ rif1Δ double mutants had a very small, but significant, increase in telomere length compared to the tel1Δ single mutant, suggesting an influence of Y' elements on telomere length regulation. We sequenced telomeres in a telomerase-null mutant (est2Δ) and found the minimal telomere length to be around 75bp. In these est2Δ mutants there were many apparent telomere recombination events at individual telomeres before the generation of survivors, and these events were significantly reduced in est2Δ rad52Δ double mutants. The rate of telomere shortening in the absence of telomerase was similar across all chromosome ends at about 5 bp per generation. This new method gives quantitative, high resolution telomere length measurement at each individual chromosome end, suggests possible new biological mechanisms regulating telomere length, and provides capability to test new hypotheses.

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Section: Telomere Shortening In the Absence Of Telomerase Reveals The Minimal Functional Telomerementioning

confidence: 99%

“…Yeast molecular cloning, culturing, sporulation, and transformation were performed as previously described in (Green and Sambrook 2012;Keener et al 2019;Shubin et al 2021).…”

Section: Molecular Cloningmentioning

confidence: 99%

Section: Molecular Cloningmentioning

confidence: 99%

Section: Yeast Strains Culturing and Transformationmentioning

confidence: 99%

Section: Southern Blotting and Densitometrymentioning

confidence: 99%

See 3 more Smart Citations

Chromosome specific telomere lengths and the minimal functional telomere revealed by nanopore sequencing

Sholes

Karimian

Gershman

et al. 2021

Preprint

Self Cite

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Tieing together loose ends: telomere instability in cancer and aging

2022

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Telomere maintenance is essential for maintaining genome integrity in both normal and cancer cells. Without functional telomeres, chromosomes lose their protective structure and undergo fusion and breakage events that drive further genome instability, including cell arrest or death. One means by which this loss can be overcome in stem cells and cancer cells is via re‐addition of G‐rich telomeric repeats by the telomerase reverse transcriptase (TERT). During aging of somatic tissues, however, insufficient telomerase expression leads to a proliferative arrest called replicative senescence, which is triggered when telomeres reach a critically short threshold that induces a DNA damage response. Cancer cells express telomerase but do not entirely escape telomere instability as they often possess short telomeres; hence there is often selection for genetic alterations in the TERT promoter that result in increased telomerase expression. In this review, we discuss our current understanding of the consequences of telomere instability in cancer and aging, and outline the opportunities and challenges that lie ahead in exploiting the reliance of cells on telomere maintenance for preserving genome stability.

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A CDK-Dependent Phosphorylation of a Novel Domain of Rif1 Regulates its Function during Telomere Damage and Other Types of Stress

Robertson

Xue

Chowdhury

et al. 2023

Molecular and Cellular Biology

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Rif1 mediates telomere length, DNA replication, and DNA damage responses in budding yeast. Previous work identified several posttranslational modifications of Rif1, however none of these was shown to mediate the molecular or cellular responses to DNA damage, including telomere damage. We searched for such modifications using immunoblotting methods and the cdc13-1 and tlc1Δ models of telomere damage. We found that Rif1 is phosphorylated during telomere damage, and that serines 57 and 110 within a novel phospho-gate domain (PGD) of Rif1 are important for this modification, in cdc13-1 cells. The phosphorylation of Rif1 appeared to inhibit its accumulation on damaged chromosomes and the proliferation of cells with telomere damage. Moreover, we found that checkpoint kinases were upstream of this Rif1 phosphorylation and that the Cdk1 activity was essential for maintaining it. Apart from telomere damage, S57 and S110 were essential for Rif1 phosphorylation during the treatment of cells with genotoxic agents or during mitotic stress. We propose a speculative “Pliers” model to explain the role of the PGD phosphorylation during telomere and other types of damage.

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Rif1 regulates telomere length through conserved HEAT repeats

Cited by 9 publications

References 44 publications

Chromosome specific telomere lengths and the minimal functional telomere revealed by nanopore sequencing

Chromosome specific telomere lengths and the minimal functional telomere revealed by nanopore sequencing

Tieing together loose ends: telomere instability in cancer and aging

A CDK-Dependent Phosphorylation of a Novel Domain of Rif1 Regulates its Function during Telomere Damage and Other Types of Stress

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