Quantitative interaction screen of telomeric repeatcontaining RNA reveals novel TERRA regulators

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Marion Scheibe - , Max Planck Institute of Biochemistry (Author)
  • Nausica Arnoult - , KU Leuven (Author)
  • Dennis Kappei - , TUD Dresden University of Technology (Author)
  • Frank Buchholz - , University Cancer Centre, Chair of Medical Systems Biology (Author)
  • Anabelle Decottignies - , KU Leuven (Author)
  • Falk Butter - , Max Planck Institute of Biochemistry (Author)
  • Matthias Mann - , Max Planck Institute of Biochemistry (Author)

Abstract

Telomeres are actively transcribed into telomeric repeat-containing RNA (TERRA), which has been implicated in the regulation of telomere length and heterochromatin formation. Here, we applied quantitative mass spectrometry (MS)-based proteomics to obtain a high-confidence interactome of TERRA. Using SILAC-labeled nuclear cell lysates in an RNA pulldown experiment and two different salt conditions, we distinguished 115 proteins binding specifically to TERRA out of a large set of background binders. While TERRA binders identified in two previous studies showed little overlap, using quantitative mass spectrometry we obtained many candidates reported in these two studies. To test whether novel candidates found here are involved in TERRA regulation, we performed an esiRNA-based interference analysis for 15 of them. Knockdown of 10 genes encoding candidate proteins significantly affected total cellular levels of TERRA, and RNAi of five candidates perturbed TERRA recruitment to telomeres. Notably, depletion of SRRT/ARS2, involved in miRNA processing, up-regulated both total and telomere-bound TERRA. Conversely, knockdown of MORF4L2, a component of the NuA4 histone acetyltransferase complex, reduced TERRA levels both globally and for telomere-bound TERRA. We thus identified new proteins involved in the homeostasis and telomeric abundance of TERRA, extending our knowledge of TERRA regulation.

Details

Original languageEnglish
Pages (from-to)2149-2157
Number of pages9
JournalGenome Research
Volume23
Issue number12
Publication statusPublished - Dec 2013
Peer-reviewedYes

External IDs

PubMed 23921659

Keywords

ASJC Scopus subject areas