TAILIEUCHUNG - Báo cáo y học: " The RNA binding protein HuR does not interact directly with HIV-1 reverse transcriptase and does not affect reverse transcription in vitro"

Tuyển tập các báo cáo nghiên cứu về y học được đăng trên tạp chí y học Retrovirology cung cấp cho các bạn kiến thức về ngành y đề tài: The RNA binding protein HuR does not interact directly with HIV-1 reverse transcriptase and does not affect reverse transcription in vitro. | Ahn etal. Retrovirology 2010 7 40 http content 7 1 40 gtr RETR0VIR0L0GY RESEARCH Open Access The RNA binding protein HuR does not interact directly with HIV-1 reverse transcriptase and does not affect reverse transcription in vitro Jinwoo Ahn 1 In-Ja L Byeon1 Sanjeewa Dharmasena2 Kelly Huber2 Jason Concel1 Angela M Gronenborn 1 and Nicolas Sluis-Cremer 2 Abstract Background Lemay etal 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 etal. 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. Background Reverse transcription of the viral single-stranded RNA genome into double-stranded DNA is a critical step in the HIV-1 life-cycle. Although the viral proteins nucle-ocapsid matrix integrase tat nef and vif may participate in the

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