Journal
NUCLEIC ACIDS RESEARCH
Volume 43, Issue 18, Pages 8884-8897Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkv897
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Funding
- Bill and Melinda Gates Foundation (GCE) [OPP1035881, OPP1097238]
- European Research Council (ERC Consolidator) [615879]
- Italian Ministry of University and Research [FIRB-Ideas RBID082ATK 001]
- National Institute of Health [GM064328]
- European Research Council (ERC) [615879] Funding Source: European Research Council (ERC)
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Folding of the LTR promoter into dynamic G-quadruplex conformations has been shown to suppress its transcriptional activity in HIV-1. Here we sought to identify the proteins that control the folding of this region of proviral genome by inducing/stabilizing G-quadruplex structures. The implementation of electrophorethic mobility shift assay and pull-down experiments coupled with mass spectrometric analysis revealed that the cellular protein nucleolin is able to specifically recognize G-quadruplex structures present in the LTR promoter. Nucleolin recognized with high affinity and specificity the majority, but not all the possible G-quadruplexes folded by this sequence. In addition, it displayed greater binding preference towards DNA than RNA G-quadruplexes, thus indicating two levels of selectivity based on the sequence and nature of the target. The interaction translated into stabilization of the LTR G-quadruplexes and increased promoter silencing activity; in contrast, disruption of nucleolin binding in cells by both siRNAs and a nucleolin binding aptamer greatly increased LTR promoter activity. These data indicate that nucleolin possesses a specific and regulated activity toward the HIV-1 LTR promoter, which ismediated by G-quadruplexes. These observations provide new essential insights into viral transcription and a possible low mutagenic target for antiretroviral therapy.
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