A dynamic i-motif with a duplex stem-loop in the long terminal repeat promoter of the HIV-1 proviral genome modulates viral transcription.


Journal

Nucleic acids research
ISSN: 1362-4962
Titre abrégé: Nucleic Acids Res
Pays: England
ID NLM: 0411011

Informations de publication

Date de publication:
02 12 2019
Historique:
accepted: 07 10 2019
revised: 27 09 2019
received: 04 07 2019
pubmed: 31 10 2019
medline: 12 5 2020
entrez: 31 10 2019
Statut: ppublish

Résumé

I-motifs are non-canonical nucleic acids structures characterized by intercalated H-bonds between hemi-protonated cytosines. Evidence on the involvement of i-motif structures in the regulation of cellular processes in human cells has been consistently growing in the recent years. However, i-motifs within non-human genomes have never been investigated. Here, we report the characterization of i-motifs within the long terminal repeat (LTR) promoter of the HIV-1 proviral genome. Biophysical and biochemical analysis revealed formation of a predominant i-motif with an unprecedented loop composition. One-dimensional nuclear magnetic resonance investigation demonstrated formation of three G-C H-bonds in the long loop, which likely improve the structure overall stability. Pull-down experiments combined with mass spectrometry and protein crosslinking analysis showed that the LTR i-motif is recognized by the cellular protein hnRNP K, which induced folding at physiological conditions. In addition, hnRNP K silencing resulted in an increased LTR promoter activity, confirming the ability of the protein to stabilize the i-motif-forming sequence, which in turn regulates the LTR-mediated HIV-1 transcription. These findings provide new insights into the complexity of the HIV-1 virus and lay the basis for innovative antiviral drug design, based on the possibility to selectively recognize and target the HIV-1 LTR i-motif.

Identifiants

pubmed: 31665504
pii: 5608988
doi: 10.1093/nar/gkz937
pmc: PMC6868428
doi:

Substances chimiques

Heterogeneous-Nuclear Ribonucleoprotein K 0
RNA, Viral 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

11057-11068

Subventions

Organisme : European Research Council
ID : 615879
Pays : International

Informations de copyright

© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.

Références

Chem Rev. 2019 May 22;119(10):6290-6325
pubmed: 30605316
EMBO J. 2002 Jul 1;21(13):3476-85
pubmed: 12093748
Nucleic Acids Res. 1996 Dec 15;24(24):5062-3
pubmed: 9016685
Biochemistry. 2014 Apr 29;53(16):2581-93
pubmed: 24735378
J Med Chem. 2013 Aug 22;56(16):6521-30
pubmed: 23865750
J Antimicrob Chemother. 2014 Dec;69(12):3248-58
pubmed: 25103489
Angew Chem Int Ed Engl. 2012 Apr 23;51(17):4067-70
pubmed: 22411471
Phys Chem Chem Phys. 2016 Mar 21;18(11):7997-8004
pubmed: 26957024
Cell Host Microbe. 2008 Oct 16;4(4):398-408
pubmed: 18854243
Hum Genet. 2016 Aug;135(8):851-67
pubmed: 27215579
Electrophoresis. 1999 Dec;20(18):3551-67
pubmed: 10612281
Chem Commun (Camb). 2015 Apr 4;51(26):5630-2
pubmed: 25686374
Chembiochem. 2017 Oct 18;18(20):2033-2044
pubmed: 28805284
PLoS Pathog. 2015 Feb 05;11(2):e1004562
pubmed: 25654363
Nucleic Acids Res. 2015 Oct 15;43(18):8884-97
pubmed: 26354862
Bioorg Med Chem. 2014 Aug 15;22(16):4407-18
pubmed: 24957878
Methods Enzymol. 1980;65(1):499-560
pubmed: 6246368
Sci Rep. 2017 Jul 18;7(1):5743
pubmed: 28720801
Sci Rep. 2017 Mar 24;7:45244
pubmed: 28338097
J Am Chem Soc. 2016 Oct 26;138(42):14138-14151
pubmed: 27669098
J Am Chem Soc. 2014 Mar 19;136(11):4161-71
pubmed: 24559410
Nucleic Acids Res. 2016 Jul 27;44(13):6442-51
pubmed: 27298260
Biochim Biophys Acta Gen Subj. 2017 May;1861(5 Pt B):1371-1381
pubmed: 27913192
Nucleic Acids Res. 2014 Nov 10;42(20):12352-66
pubmed: 25332402
J Am Chem Soc. 2014 Mar 19;136(11):4172-85
pubmed: 24559432
Anal Chim Acta. 2016 Aug 3;930:1-12
pubmed: 27265899
Cell Chem Biol. 2019 Aug 15;26(8):1110-1121.e4
pubmed: 31155510
Nature. 1993 Jun 10;363(6429):561-5
pubmed: 8389423
Biophys J. 2003 Jun;84(6):3838-47
pubmed: 12770889
Nucleic Acids Res. 2018 Apr 20;46(7):3270-3283
pubmed: 29554280
Nucleic Acids Res. 2018 Sep 19;46(16):8038-8056
pubmed: 30124962
J Am Chem Soc. 2017 Jun 28;139(25):8522-8536
pubmed: 28570076
Curr Top Med Chem. 2015;15(19):1940-6
pubmed: 25980416
Chem Soc Rev. 2008 Jul;37(7):1375-84
pubmed: 18568163
J Med Chem. 2015 Dec 24;58(24):9639-52
pubmed: 26599611
Biochemistry. 2014 Mar 18;53(10):1586-94
pubmed: 24564458
Nucleic Acids Res. 2002 Nov 1;30(21):4618-25
pubmed: 12409451
Cell Biochem Funct. 2016 Jun;34(4):191-6
pubmed: 27001679
Nat Chem. 2018 Jun;10(6):631-637
pubmed: 29686376
J Am Chem Soc. 2018 Oct 24;140(42):13654-13662
pubmed: 30299955
J Am Chem Soc. 2014 Apr 9;136(14):5249-52
pubmed: 24649937
Nucleic Acids Res. 2015 Oct 15;43(18):8627-37
pubmed: 26350216
Molecules. 2013 Oct 08;18(10):12368-95
pubmed: 24108400
J Virol. 1996 Oct;70(10):7049-55
pubmed: 8794349
Annu Rev Microbiol. 2018 Sep 08;72:49-69
pubmed: 29852085
Sci Rep. 2017 May 17;7(1):2018
pubmed: 28515481

Auteurs

Emanuela Ruggiero (E)

Department of Molecular Medicine, University of Padua, 35121 Padua, Italy.

Sara Lago (S)

Department of Molecular Medicine, University of Padua, 35121 Padua, Italy.

Primož Šket (P)

Slovenian NMR center, National Institute of Chemistry, Hajdrihova, 19, Ljubljana SI-1000, Slovenia.

Matteo Nadai (M)

Department of Molecular Medicine, University of Padua, 35121 Padua, Italy.

Ilaria Frasson (I)

Department of Molecular Medicine, University of Padua, 35121 Padua, Italy.

Janez Plavec (J)

Slovenian NMR center, National Institute of Chemistry, Hajdrihova, 19, Ljubljana SI-1000, Slovenia.

Sara N Richter (SN)

Department of Molecular Medicine, University of Padua, 35121 Padua, Italy.

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