Posttranscriptional manipulation of TERC reverses molecular hallmarks of telomere disease
Baris Boyraz, … , Patrick Cahan, Suneet Agarwal
Baris Boyraz, … , Patrick Cahan, Suneet Agarwal
Published August 2, 2016
Citation Information: J Clin Invest. 2016;126(9):3377-3382. https://doi.org/10.1172/JCI87547.
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Brief Report Genetics

Posttranscriptional manipulation of TERC reverses molecular hallmarks of telomere disease

Abstract

The telomerase RNA component (TERC) is a critical determinant of cellular self-renewal. Poly(A)-specific ribonuclease (PARN) is required for posttranscriptional maturation of TERC. PARN mutations lead to incomplete 3′ end processing and increased destruction of nascent TERC RNA transcripts, resulting in telomerase deficiency and telomere diseases. Here, we determined that overexpression of TERC increased telomere length in PARN-deficient cells and hypothesized that decreasing posttranscriptional 3′ oligo-adenylation of TERC would counteract the deleterious effects of PARN mutations. Inhibition of the noncanonical poly(A) polymerase PAP-associated domain–containing 5 (PAPD5) increased TERC levels in PARN-mutant patient cells. PAPD5 inhibition was also associated with increases in TERC stability, telomerase activity, and telomere elongation. Our results demonstrate that manipulating posttranscriptional regulatory pathways may be a potential strategy to reverse the molecular hallmarks of telomere disease.

Authors

Baris Boyraz, Diane H. Moon, Matthew Segal, Maud Z. Muosieyiri, Asli Aykanat, Albert K. Tai, Patrick Cahan, Suneet Agarwal

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Figure 3

PAPD5 inhibition rescues TERC maturation, telomerase activity, and telomere length in PARN-mutant patient cells.

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PAPD5 inhibition rescues TERC maturation, telomerase activity, and telo...
(A) 3′ RACE TERC amplicons from normal (WT) versus PARN-mutant iPSCs after lentiviral shRNA directed against PAPD5 versus luciferase (ctrl). (B) Representative Northern blot of TERC RNA from WT versus PARN-mutant iPSCs after lentiviral shRNA directed against PAPD5 versus luciferase. (C) Relative TERC levels from Northern blots (B) are quantified relative to WT cells with control knockdown *P ≤ 0.05; NS: not significant. n = 3. (D) Representative Northern blot of TERC levels in WT and PARN-mutant iPSCs after lentiviral shRNA directed against PAPD5 versus luciferase at 0–4 hours following actinomycin treatment. TERC levels are normalized relative to time 0. (E) Graph of TERC decay rate. Dashed line reflects slope determined by simple linear regression. (F) TRAP assay in iPSCs after lentiviral shRNA against PAPD5 versus luciferase. n = 3. (G) TRF of WT and PARN-mutant patient iPSCs after lentiviral shRNA against PAPD5 versus luciferase over time in culture.