The RNA binding protein HuR does not interact directly with HIV-1 reverse transcriptase and does not affect reverse transcription in vitro

• Main Authors: Gronenborn Angela M, Concel Jason, Huber Kelly, Dharmasena Sanjeewa, Byeon In-Ja L, Ahn Jinwoo, Sluis-Cremer Nicolas
• Format: Article
• Language: English
• Published: BMC 2010-05-01
• Series: Retrovirology
• Online Access: http://www.retrovirology.com/content/7/1/40

<p>Abstract</p> <p>Background</p> <p>Lemay <it>et al </it>recently reported that the RNA binding protein HuR directly interacts with the ribonuclease H (RNase H) domain of HIV-1 reverse transcriptase (RT) and influences the efficiency of viral reverse transcription (Lemay <it>et al</it>., 2008, Retrovirology 5:47). HuR is a member of the embryonic lethal abnormal vision protein family and contains 3 RNA recognition motifs (RRMs) that bind AU-rich elements (AREs). To define the structural determinants of the HuR-RT interaction and to elucidate the mechanism(s) by which HuR influences HIV-1 reverse transcription activity <it>in vitro</it>, we cloned and purified full-length HuR as well as three additional protein constructs that contained the N-terminal and internal RRMs, the internal and C-terminal RRMs, or the C-terminal RRM only.</p> <p>Results</p> <p>All four HuR proteins were purified and characterized by biophysical methods. They are well structured and exist as monomers in solution. No direct protein-protein interaction between HuR and HIV-1 RT was detected using NMR titrations with ¹⁵N labeled HuR variants or the ¹⁵N labeled RNase H domain of HIV-1 RT. Furthermore, HuR did not significantly affect the kinetics of HIV-1 reverse transcription <it>in vitro</it>, even on RNA templates that contain AREs.</p> <p>Conclusions</p> <p>Our results suggest that HuR does not impact HIV-1 replication through a direct protein-protein interaction with the viral RT.</p>