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

abstract

  • BACKGROUND: Lemay et al 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 et al., 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 in vitro, 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. RESULTS: 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 in vitro, even on RNA templates that contain AREs. CONCLUSIONS: Our results suggest that HuR does not impact HIV-1 replication through a direct protein-protein interaction with the viral RT.

published proceedings

  • Retrovirology

author list (cited authors)

  • Ahn, J., Byeon, I., Dharmasena, S., Huber, K., Concel, J., Gronenborn, A. M., & Sluis-Cremer, N.

citation count

  • 8

complete list of authors

  • Ahn, Jinwoo||Byeon, In-Ja L||Dharmasena, Sanjeewa||Huber, Kelly||Concel, Jason||Gronenborn, Angela M||Sluis-Cremer, Nicolas

publication date

  • January 2010