PMID- 11504876
OWN - NLM
STAT- MEDLINE
DCOM- 20010906
LR  - 20190501
IS  - 1362-4962 (Electronic)
IS  - 0305-1048 (Print)
IS  - 0305-1048 (Linking)
VI  - 29
IP  - 16
DP  - 2001 Aug 15
TI  - Human telomerase RNA-protein interactions.
PG  - 3385-93
AB  - Telomere length is maintained in most eukaryotic cells by telomerase. The core 
      components of this ribonucleoprotein (RNP) enzyme include a protein catalytic 
      subunit, composed of motifs conserved among reverse transcriptases (RT), and an 
      RNA subunit that contains a short template sequence essential for the synthesis 
      of telomeric repeats. We developed an electrophoretic mobility shift assay using 
      active telomerase partially purified from 293 cells and radiolabeled, in 
      vitro-transcribed human telomerase RNA (hTR) to investigate the molecular 
      interactions of the human telomerase RT (hTERT) and telomerase-associated 
      proteins with hTR. A specific hTR-protein complex was identified and shown to 
      contain hTERT and human Staufen by antibody supershift assays. Variants of hTR 
      altered in distinct structural elements were analyzed for their ability to 
      competitively inhibit complex formation. Human telomerase RNAs lacking the 
      CR4-CR5 domain were poor inhibitors of hTR-protein complex formation, suggesting 
      that the CR4-CR5 domain of hTR is a potential protein-binding site. Furthermore, 
      alterations in the telomerase RNA pseudoknot's P3 helix, the CR7 domain, or the 
      H/ACA box efficiently inhibited formation of the complex, indicating that these 
      domains are dispensable for the assembly of a telomerase RNP in vitro. Potential 
      telomerase-associated proteins that bind hTR were also identified using a UV 
      cross-linking assay.
FAU - Bachand, F
AU  - Bachand F
AD  - Department of Anatomy and Cell Biology, McGill University, Montreal, Quebec H3A 
      2B2, Canada.
FAU - Triki, I
AU  - Triki I
FAU - Autexier, C
AU  - Autexier C
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Nucleic Acids Res
JT  - Nucleic acids research
JID - 0411011
RN  - 0 (Cell Extracts)
RN  - 0 (Cytoskeletal Proteins)
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (RNA-Binding Proteins)
RN  - 0 (STAU1 protein, human)
RN  - 0 (staufen protein, mammalian)
RN  - 0 (telomerase RNA)
RN  - 63231-63-0 (RNA)
RN  - EC 2.7.7.49 (Telomerase)
SB  - IM
MH  - Binding Sites
MH  - Binding, Competitive
MH  - Catalysis
MH  - Catalytic Domain
MH  - Cell Extracts
MH  - Cell Line, Transformed
MH  - Cytoskeletal Proteins
MH  - DNA-Binding Proteins
MH  - Humans
MH  - Inhibitory Concentration 50
MH  - Molecular Weight
MH  - Mutation/genetics
MH  - Nucleic Acid Conformation
MH  - RNA/chemistry/genetics/*metabolism
MH  - RNA-Binding Proteins/antagonists & inhibitors/chemistry/genetics/*metabolism
MH  - Telomerase/antagonists & inhibitors/chemistry/genetics/*metabolism
MH  - Ultraviolet Rays
PMC - PMC55859
EDAT- 2001/08/16 10:00
MHDA- 2001/09/08 10:01
PMCR- 2001/08/15
CRDT- 2001/08/16 10:00
PHST- 2001/08/16 10:00 [pubmed]
PHST- 2001/09/08 10:01 [medline]
PHST- 2001/08/16 10:00 [entrez]
PHST- 2001/08/15 00:00 [pmc-release]
AID - gke476 [pii]
AID - 10.1093/nar/29.16.3385 [doi]
PST - ppublish
SO  - Nucleic Acids Res. 2001 Aug 15;29(16):3385-93. doi: 10.1093/nar/29.16.3385.