Rif1 acts through Protein Phosphatase 1 but independent of replication timing to suppress telomere extension in budding yeast

Sylwia Kedziora, Vamsi K Gali, Rosemary H C Wilson, Kate R M Clark, Conrad A Nieduszynski, Shin-ichiro Hiraga, Anne D Donaldson (Corresponding Author)

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Abstract

The Rif1 protein negatively regulates telomeric TG repeat length in the budding yeast Saccharomyces cerevisiae, but how it prevents telomere over-extension is unknown. Rif1 was recently shown to control DNA replication by acting as a Protein Phosphatase 1 (PP1)-targeting subunit. Therefore, we investigated whether Rif1 controls telomere length by targeting PP1 activity. We find that a Rif1 mutant defective for PP1 interaction causes a long-telomere phenotype, similar to that of rif1Δ cells. Tethering PP1 at a specific telomere partially substitutes for Rif1 in limiting TG repeat length, confirming the importance of PP1 in telomere length control. Ablating Rif1–PP1 interaction is known to cause precocious activation of telomere-proximal replication origins and aberrantly early telomere replication. However, we find that Rif1 still limits telomere length even if late replication is forced through deletion of nearby replication origins, indicating that Rif1 can control telomere length independent of replication timing. Moreover we find that, even at a de novo telomere created after DNA synthesis during a mitotic block, Rif1–PP1 interaction is required to suppress telomere lengthening and prevent inappropriate recruitment of Tel1 kinase. Overall, our results show that Rif1 controls telomere length by recruiting PP1 to directly suppress telomerase-mediated TG repeat lengthening.
Original languageEnglish
Pages (from-to)3993-4003
Number of pages11
JournalNucleic Acids Research
Volume46
Issue number8
Early online date26 Feb 2018
DOIs
Publication statusPublished - 4 May 2018

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Protein Phosphatase 1
Saccharomycetales
Telomere
Replication Origin
Telomere Homeostasis
Telomerase
Mutant Proteins
DNA Replication
Saccharomyces cerevisiae
Phosphotransferases

Keywords

  • telomere
  • chromosome
  • Rif1
  • phosphatase
  • replication timing

Cite this

Rif1 acts through Protein Phosphatase 1 but independent of replication timing to suppress telomere extension in budding yeast. / Kedziora, Sylwia; Gali, Vamsi K; Wilson, Rosemary H C; Clark, Kate R M; Nieduszynski, Conrad A; Hiraga, Shin-ichiro; Donaldson, Anne D (Corresponding Author).

In: Nucleic Acids Research, Vol. 46, No. 8, 04.05.2018, p. 3993-4003.

Research output: Contribution to journalArticle

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title = "Rif1 acts through Protein Phosphatase 1 but independent of replication timing to suppress telomere extension in budding yeast",
abstract = "The Rif1 protein negatively regulates telomeric TG repeat length in the budding yeast Saccharomyces cerevisiae, but how it prevents telomere over-extension is unknown. Rif1 was recently shown to control DNA replication by acting as a Protein Phosphatase 1 (PP1)-targeting subunit. Therefore, we investigated whether Rif1 controls telomere length by targeting PP1 activity. We find that a Rif1 mutant defective for PP1 interaction causes a long-telomere phenotype, similar to that of rif1Δ cells. Tethering PP1 at a specific telomere partially substitutes for Rif1 in limiting TG repeat length, confirming the importance of PP1 in telomere length control. Ablating Rif1–PP1 interaction is known to cause precocious activation of telomere-proximal replication origins and aberrantly early telomere replication. However, we find that Rif1 still limits telomere length even if late replication is forced through deletion of nearby replication origins, indicating that Rif1 can control telomere length independent of replication timing. Moreover we find that, even at a de novo telomere created after DNA synthesis during a mitotic block, Rif1–PP1 interaction is required to suppress telomere lengthening and prevent inappropriate recruitment of Tel1 kinase. Overall, our results show that Rif1 controls telomere length by recruiting PP1 to directly suppress telomerase-mediated TG repeat lengthening.",
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author = "Sylwia Kedziora and Gali, {Vamsi K} and Wilson, {Rosemary H C} and Clark, {Kate R M} and Nieduszynski, {Conrad A} and Shin-ichiro Hiraga and Donaldson, {Anne D}",
note = "ACKNOWLEDGEMENTS We thank Katsunori Sugimoto for strains, plasmids and technical advice. We thank Amanda Williams for Illumina NextSeq 500 technical support and advice. Takashi Kubota provided helpful comments on the manuscript. FUNDING Cancer Research UK Programme Award [A19059 to A.D., S.H.]; Wellcome Trust Investigator Award [110064/Z/15/Z to C.A.N., R.H.C.W.]; SULSA PhD Studentship (to S.K.). Funding for open access charge: Cancer Research UK via University of Aberdeen Open Access Fund. Conflict of interest statement. None declared.",
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T1 - Rif1 acts through Protein Phosphatase 1 but independent of replication timing to suppress telomere extension in budding yeast

AU - Kedziora, Sylwia

AU - Gali, Vamsi K

AU - Wilson, Rosemary H C

AU - Clark, Kate R M

AU - Nieduszynski, Conrad A

AU - Hiraga, Shin-ichiro

AU - Donaldson, Anne D

N1 - ACKNOWLEDGEMENTS We thank Katsunori Sugimoto for strains, plasmids and technical advice. We thank Amanda Williams for Illumina NextSeq 500 technical support and advice. Takashi Kubota provided helpful comments on the manuscript. FUNDING Cancer Research UK Programme Award [A19059 to A.D., S.H.]; Wellcome Trust Investigator Award [110064/Z/15/Z to C.A.N., R.H.C.W.]; SULSA PhD Studentship (to S.K.). Funding for open access charge: Cancer Research UK via University of Aberdeen Open Access Fund. Conflict of interest statement. None declared.

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N2 - The Rif1 protein negatively regulates telomeric TG repeat length in the budding yeast Saccharomyces cerevisiae, but how it prevents telomere over-extension is unknown. Rif1 was recently shown to control DNA replication by acting as a Protein Phosphatase 1 (PP1)-targeting subunit. Therefore, we investigated whether Rif1 controls telomere length by targeting PP1 activity. We find that a Rif1 mutant defective for PP1 interaction causes a long-telomere phenotype, similar to that of rif1Δ cells. Tethering PP1 at a specific telomere partially substitutes for Rif1 in limiting TG repeat length, confirming the importance of PP1 in telomere length control. Ablating Rif1–PP1 interaction is known to cause precocious activation of telomere-proximal replication origins and aberrantly early telomere replication. However, we find that Rif1 still limits telomere length even if late replication is forced through deletion of nearby replication origins, indicating that Rif1 can control telomere length independent of replication timing. Moreover we find that, even at a de novo telomere created after DNA synthesis during a mitotic block, Rif1–PP1 interaction is required to suppress telomere lengthening and prevent inappropriate recruitment of Tel1 kinase. Overall, our results show that Rif1 controls telomere length by recruiting PP1 to directly suppress telomerase-mediated TG repeat lengthening.

AB - The Rif1 protein negatively regulates telomeric TG repeat length in the budding yeast Saccharomyces cerevisiae, but how it prevents telomere over-extension is unknown. Rif1 was recently shown to control DNA replication by acting as a Protein Phosphatase 1 (PP1)-targeting subunit. Therefore, we investigated whether Rif1 controls telomere length by targeting PP1 activity. We find that a Rif1 mutant defective for PP1 interaction causes a long-telomere phenotype, similar to that of rif1Δ cells. Tethering PP1 at a specific telomere partially substitutes for Rif1 in limiting TG repeat length, confirming the importance of PP1 in telomere length control. Ablating Rif1–PP1 interaction is known to cause precocious activation of telomere-proximal replication origins and aberrantly early telomere replication. However, we find that Rif1 still limits telomere length even if late replication is forced through deletion of nearby replication origins, indicating that Rif1 can control telomere length independent of replication timing. Moreover we find that, even at a de novo telomere created after DNA synthesis during a mitotic block, Rif1–PP1 interaction is required to suppress telomere lengthening and prevent inappropriate recruitment of Tel1 kinase. Overall, our results show that Rif1 controls telomere length by recruiting PP1 to directly suppress telomerase-mediated TG repeat lengthening.

KW - telomere

KW - chromosome

KW - Rif1

KW - phosphatase

KW - replication timing

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