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HIV-1 Pr55(Gag) binds genomic and spliced RNAs with different affinity and stoichiometry

Bernacchi, Serena, Abd El-Wahab, Ekram W., Dubois, Noe, Hijnen, Marcel, Smyth, Redmond P., Mak, Johnson, Marquet, Roland and Paillart, Jean-Christophe 2017, HIV-1 Pr55(Gag) binds genomic and spliced RNAs with different affinity and stoichiometry, RNA biology, vol. 14, no. 1, pp. 90-103, doi: 10.1080/15476286.2016.1256533.

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Title HIV-1 Pr55(Gag) binds genomic and spliced RNAs with different affinity and stoichiometry
Author(s) Bernacchi, Serena
Abd El-Wahab, Ekram W.
Dubois, Noe
Hijnen, Marcel
Smyth, Redmond P.
Mak, Johnson
Marquet, Roland
Paillart, Jean-Christophe
Journal name RNA biology
Volume number 14
Issue number 1
Start page 90
End page 103
Total pages 15
Publisher Taylor & Francis
Place of publication Abingdon, Eng.
Publication date 2017-01
ISSN 1547-6286
1555-8584
Keyword(s) Fluorescence spectroscopy
HIV-1
Pr55Gag
genomic RNA selection
high affinity binding site
protein-RNA interaction
stoichiometry
Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
Pr55(Gag)
HUMAN-IMMUNODEFICIENCY-VIRUS
DIMERIZATION INITIATION SITE
ACID CHAPERONE ACTIVITIES
RETROVIRAL GAG PROTEINS
KISSING-LOOP HAIRPIN
IN-VITRO EVIDENCE
NUCLEOCAPSID PROTEIN
TYPE-1 RNA
REVERSE TRANSCRIPTION
PACKAGING SIGNAL
Summary The HIV-1 Pr55(Gag) precursor specifically selects genomic RNA (gRNA) from a large variety of cellular and spliced viral RNAs (svRNAs), however the molecular mechanisms of this selective recognition remains poorly understood. To gain better understanding of this process, we analyzed the interactions between Pr55(Gag) and a large panel of viral RNA (vRNA) fragments encompassing the main packaging signal (Psi) and its flanking regions by fluorescence spectroscopy. We showed that the gRNA harbors a high affinity binding site which is absent from svRNA species, suggesting that this site might be crucial for selecting the HIV-1 genome. Our stoichiometry analysis of protein/RNA complexes revealed that few copies of Pr55(Gag) specifically associate with the 5' region of the gRNA. Besides, we found that gRNA dimerization significantly impacts Pr55(Gag) binding, and we confirmed that the internal loop of stem-loop 1 (SL1) in Psi is crucial for specific interaction with Pr55(Gag). Our analysis of gRNA fragments of different length supports the existence of a long-range tertiary interaction involving sequences upstream and downstream of the Psi region. This long-range interaction might promote optimal exposure of SL1 for efficient Pr55(Gag) recognition. Altogether, our results shed light on the molecular mechanisms allowing the specific selection of gRNA by Pr55(Gag) among a variety of svRNAs, all harboring SL1 in their first common exon.
Language eng
DOI 10.1080/15476286.2016.1256533
Field of Research 110801 Medical Bacteriology
0604 Genetics
Socio Economic Objective 920499 Public Health (excl. Specific Population Health) not elsewhere classified
HERDC Research category C1 Refereed article in a scholarly journal
Copyright notice ©2017, Taylor & Francis Group, LLC
Persistent URL http://hdl.handle.net/10536/DRO/DU:30091593

Document type: Journal Article
Collection: School of Medicine
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