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PMID: 25907625 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Targeted Detection of G-Quadruplexes in Cellular RNAs.

Angewandte Chemie (International ed. in English) ·Vol. 54 ·No. 23 ·2015-06-01 ·Pages 6751-4

Kwok CK, Balasubramanian S

Abstract

The G-quadruplex (G4) is a non-canonical nucleic acid structure which regulates important cellular processes. RNA G4s have recently been shown to exist in human cells and be biologically significant. Described herein is a new approach to detect and map RNA G4s in cellular transcripts. This method exploits the specific control of RNA G4-cation and RNA G4-ligand interactions during reverse transcription, by using a selective reverse transcriptase to monitor RNA G4-mediated reverse transcriptase stalling (RTS) events. Importantly, a ligation-amplification strategy is coupled with RTS, and enables detection and mapping of G4s in important, low-abundance cellular RNAs. Strong evidence is provided for G4 formation in full-length cellular human telomerase RNA, offering important insights into its cellular function.

Keywords
G-quadruplexes RNA structures cations ligand effects reverse transcription telomerase RNA
MeSH Terms
G-Quadruplexes HeLa Cells Humans Molecular Structure RNA/chemistry,metabolism Telomerase/chemistry,metabolism
Chemicals
telomerase RNA RNA Telomerase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Kwok Chun Kit
The University of Cambridge, Department of Chemistry, Lensfield Road, Cambridge, CB2 1EW (UK).
Balasubramanian Shankar
The University of Cambridge, Department of Chemistry, Lensfield Road, Cambridge, CB2 1EW (UK). sb10031@cam.ac.uk.
References (40)
40 references, click to expand
  1. In-line probing of RNA G-quadruplexes.
    Methods. 2013 Nov;64(1):79-87 PMID: 23500045
  2. Determination of in vivo RNA structure in low-abundance transcripts.
    Nat Commun. 2013;4:2971 PMID: 24336128
  3. G-quadruplexes in promoters throughout the human genome.
    Nucleic Acids Res. 2007;35(2):406-13 PMID: 17169996
  4. QGRS Mapper: a web-based server for predicting G-quadruplexes in nucleotide sequences.
    Nucleic Acids Res. 2006 Jul 1;34(Web Server issue):W676-82 PMID: 16845096
  5. Effects of cationic porphyrins as G-quadruplex interactive agents in human tumor cells.
    Cancer Res. 1999 Feb 1;59(3):639-44 PMID: 9973212
  6. Elevated levels of G-quadruplex formation in human stomach and liver cancer tissues.
    PLoS One. 2014;9(7):e102711 PMID: 25033211
  7. Following G-quartet formation by UV-spectroscopy.
    FEBS Lett. 1998 Sep 11;435(1):74-8 PMID: 9755862
  8. The selectivity for K+ versus Na+ in DNA quadruplexes is dominated by relative free energies of hydration: a thermodynamic analysis by 1H NMR.
    Biochemistry. 1996 Dec 3;35(48):15383-90 PMID: 8952490
  9. Expression of telomerase RNA template, but not telomerase reverse transcriptase, is limiting for telomere length maintenance in vivo.
    Mol Cell Biol. 2004 Aug;24(16):7024-31 PMID: 15282303
  10. Template boundary definition in mammalian telomerase.
    Genes Dev. 2003 Nov 15;17(22):2747-52 PMID: 14630939
  11. Highly efficient G-quadruplex recognition by bisquinolinium compounds.
    J Am Chem Soc. 2007 Feb 21;129(7):1856-7 PMID: 17260991
  12. An RNA G-quadruplex in the 5' UTR of the NRAS proto-oncogene modulates translation.
    Nat Chem Biol. 2007 Apr;3(4):218-21 PMID: 17322877
  13. NMR methods for studying quadruplex nucleic acids.
    Methods. 2007 Dec;43(4):264-77 PMID: 17967697
  14. G-Quadruplex formation interferes with P1 helix formation in the RNA component of telomerase hTERC.
    Chembiochem. 2008 Sep 1;9(13):2075-9 PMID: 18683270
  15. G-quadruplexes: the beginning and end of UTRs.
    Nucleic Acids Res. 2008 Nov;36(19):6260-8 PMID: 18832370
  16. A novel small molecule that alters shelterin integrity and triggers a DNA-damage response at telomeres.
    J Am Chem Soc. 2008 Nov 26;130(47):15758-9 PMID: 18975896
  17. An unusually stable G-quadruplex within the 5'-UTR of the MT3 matrix metalloproteinase mRNA represses translation in eukaryotic cells.
    Biochemistry. 2009 Jun 16;48(23):5313-9 PMID: 19397366
  18. G-quadruplex structures: in vivo evidence and function.
    Trends Cell Biol. 2009 Aug;19(8):414-22 PMID: 19589679
  19. Overview of formation of G-quadruplex structures.
    Curr Protoc Nucleic Acid Chem. 2010 Mar;Chapter 17:Unit 17.2.1-17 PMID: 20201027
  20. A reverse transcriptase stop assay revealed diverse quadruplex formations in UTRs in mRNA.
    Bioorg Med Chem Lett. 2010 Apr 1;20(7):2350-3 PMID: 20206514
  21. The BCL-2 5' untranslated region contains an RNA G-quadruplex-forming motif that modulates protein expression.
    Biochemistry. 2010 Sep 28;49(38):8300-6 PMID: 20726580
  22. A G-quadruplex structure within the 5'-UTR of TRF2 mRNA represses translation in human cells.
    Nucleic Acids Res. 2010 Nov;38(20):7187-98 PMID: 20571083
  23. The 5' guanosine tracts of human telomerase RNA are recognized by the G-quadruplex binding domain of the RNA helicase DHX36 and function to increase RNA accumulation.
    Mol Cell Biol. 2011 Feb;31(4):736-43 PMID: 21149580
  24. The DEAH-box RNA helicase RHAU binds an intramolecular RNA G-quadruplex in TERC and associates with telomerase holoenzyme.
    Nucleic Acids Res. 2011 Nov;39(21):9390-404 PMID: 21846770
  25. Visualization and selective chemical targeting of RNA G-quadruplex structures in the cytoplasm of human cells.
    Nat Chem. 2014 Jan;6(1):75-80 PMID: 24345950
  26. Detection of G-quadruplex DNA in mammalian cells.
    Nucleic Acids Res. 2014 Jan;42(2):860-9 PMID: 24163102
  27. Formation of a stacked dimeric G-quadruplex containing bulges by the 5'-terminal region of human telomerase RNA (hTERC).
    Biochemistry. 2014 Mar 18;53(10):1595-600 PMID: 24601523
  28. Existence and consequences of G-quadruplex structures in DNA.
    Curr Opin Genet Dev. 2014 Apr;25:22-9 PMID: 24584093
  29. RNA G-quadruplexes cause eIF4A-dependent oncogene translation in cancer.
    Nature. 2014 Sep 4;513(7516):65-70 PMID: 25079319
  30. Dual binding of an antibody and a small molecule increases the stability of TERRA G-quadruplex.
    Angew Chem Int Ed Engl. 2015 Jan 12;54(3):910-3 PMID: 25421962
  31. A stable RNA G-quadruplex within the 5'-UTR of Arabidopsis thaliana ATR mRNA inhibits translation.
    Biochem J. 2015 Apr 1;467(1):91-102 PMID: 25793418
  32. Inventory of telomerase components in human cells reveals multiple subpopulations of hTR and hTERT.
    Nucleic Acids Res. 2014 Jul;42(13):8565-77 PMID: 24990373
  33. Effect of loop sequence and loop length on the intrinsic fluorescence of G-quadruplexes.
    Biochemistry. 2013 May 7;52(18):3019-21 PMID: 23621657
  34. A hybridization-based approach for quantitative and low-bias single-stranded DNA ligation.
    Anal Biochem. 2013 Apr 15;435(2):181-6 PMID: 23399535
  35. Quantitative visualization of DNA G-quadruplex structures in human cells.
    Nat Chem. 2013 Mar;5(3):182-6 PMID: 23422559
  36. Decrease in RNA folding cooperativity by deliberate population of intermediates in RNA G-quadruplexes.
    Angew Chem Int Ed Engl. 2013 Jan 7;52(2):683-6 PMID: 23166020
  37. Selective RNA versus DNA G-quadruplex targeting by in situ click chemistry.
    Angew Chem Int Ed Engl. 2012 Oct 29;51(44):11073-8 PMID: 23038154
  38. Circular dichroism and guanine quadruplexes.
    Methods. 2012 May;57(1):64-75 PMID: 22450044
  39. 5'-UTR RNA G-quadruplexes: translation regulation and targeting.
    Nucleic Acids Res. 2012 Jun;40(11):4727-41 PMID: 22351747
  40. The RNA helicase RHAU (DHX36) unwinds a G4-quadruplex in human telomerase RNA and promotes the formation of the P1 helix template boundary.
    Nucleic Acids Res. 2012 May;40(9):4110-24 PMID: 22238380
Article Info
Journal
Angewandte Chemie (International ed. in English)
Abbr.
Angew Chem Int Ed Engl
ISSN
1521-3773
DOI
10.1002/anie.201500891
Email
sb10031@cam.ac.uk
Published
2015-06-01
Epub
2015-00-23
Pages
6751-4
Language
English
Region
Germany
NLM ID
0370543
PMCID
PMC4510783
Subset
IM
External Links
PubMed source DOI: 10.1002/anie.201500891 PMC Full Text View journal details
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