Functional Interactions of Kluyveromyces lactis Telomerase Reverse Transcriptase with the Three-Way Junction and the Template Domains of Telomerase RNA

Wasif Al-Shareef, Yogev Brown, Christopher Bryan, Elena Shuvaeva, Shhadeh Bsoul, Raanan Greenman, Majdi M. Kabaha, Nikolai B. Ulyanov, Emmanuel Skordalakes*, Yehuda Tzfati*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Scopus citations

Abstract

The ribonucleoprotein telomerase contains two essential components: telomerase RNA (TER) and telomerase reverse transcriptase (TERT, Est2 in yeast). A small portion of TER, termed the template, is copied by TERT onto the chromosome ends, thus compensating for sequence loss due to incomplete DNA replication and nuclease action. Although telomerase RNA is highly divergent in sequence and length across fungi and mammals, structural motifs essential for telomerase function are conserved. Here, we show that Est2 from the budding yeast Kluyveromyces lactis (klEst2) binds specifically to an essential three-way junction (TWJ) structure in K. lactis TER, which shares a conserved structure and sequence features with the essential CR4-CR5 domain of vertebrate telomerase RNA. klEst2 also binds specifically to the template domain, independently and mutually exclusive of its interaction with TWJ. Furthermore, we present the high-resolution structure of the klEst2 telomerase RNA-binding domain (klTRBD). Mutations introduced in vivo in klTRBD based on the solved structure or in TWJ based on its predicted RNA structure caused severe telomere shortening. These results demonstrate the conservation and importance of these domains and the multiple protein–RNA interactions between Est2 and TER for telomerase function.

Original languageAmerican English
Article number10757
JournalInternational Journal of Molecular Sciences
Volume23
Issue number18
DOIs
StatePublished - Sep 2022

Bibliographical note

Publisher Copyright:
© 2022 by the authors.

Keywords

  • Est2
  • crystal structure
  • ribonucleoprotein
  • telomerase
  • telomerase RNA
  • three-way junction
  • yeast

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