A critical stem-loop structure in the CR4-CR5 domain of mammalian telomerase RNA

Julian Chen, Kay Keyer Opperman, Carol W. Greider

Research output: Contribution to journalArticle

110 Citations (Scopus)

Abstract

Telomerase is an enzyme that maintains telomere length by adding telomeric sequence repeats onto chromosome ends. The telomerase ribonucleo-protein complex consists of two essential components, a reverse transcriptase and an RNA molecule that provides the template for telomeric repeat synthesis. A common secondary structure of vertebrate telomerase RNA has been proposed based on a phylogenetic comparative analysis of 35 sequences. Here we report the identification of an additional essential base-paired region in the CR4-CR5 domain of mammalian telomerase RNA, termed P6.1. Mouse telomerase RNAs with mutations that disrupted base pairings in the P6.1 helix were unable to reconstitute telomerase activity in vivo. In contrast, an RNA mutant with compensatory mutations that restored base pairings in the P6.1 helix restored telomerase activity. In an in vitro reconstitution system stable base pairing of the P6.1 stem was required for the RNA-protein interaction between the CR4-CR5 domain and the telomerase reverse transcriptase (TERT) protein. Interestingly, two RNA mutations, one that extends the P6.1 stem and one that alters the conserved nucleotides of the L6.1 loop, allowed RNA-protein binding but significantly impaired telomerase activity. These data establish the presence of the P6.1 stem-loop and its importance for the assembly and enzymatic activity of the mammalian telomerase complex.

Original languageEnglish (US)
Pages (from-to)592-597
Number of pages6
JournalNucleic Acids Research
Volume30
Issue number2
StatePublished - Jan 15 2002
Externally publishedYes

Fingerprint

Telomerase
Base Pairing
RNA
Mutation
Proteins
RNA-Binding Proteins
RNA-Directed DNA Polymerase
Telomere
telomerase RNA
Sequence Analysis
Vertebrates
Nucleotides
Chromosomes
Enzymes

ASJC Scopus subject areas

  • Genetics

Cite this

A critical stem-loop structure in the CR4-CR5 domain of mammalian telomerase RNA. / Chen, Julian; Opperman, Kay Keyer; Greider, Carol W.

In: Nucleic Acids Research, Vol. 30, No. 2, 15.01.2002, p. 592-597.

Research output: Contribution to journalArticle

Chen, Julian ; Opperman, Kay Keyer ; Greider, Carol W. / A critical stem-loop structure in the CR4-CR5 domain of mammalian telomerase RNA. In: Nucleic Acids Research. 2002 ; Vol. 30, No. 2. pp. 592-597.
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