Anticodon-like binding of the HIV-1 tRNA-like element to human lysyl-tRNA synthetase

作者：Liu Sheng; Comandur Roopa; Jones Christopher P; Tsang Pearl^*; Musier Forsyth Karin^*

来源：RNA-A Publication of the RNA Society, 2016, 22(12): 1828-1835.

DOI：10.1261/rna.058081.116

• anticodon-binding domain
• human lysyl-tRNA synthetase
• tRNA-like element
• HIV-1 genome
• NMR
• chemical shift perturbation

A critical step in the HIV-1 lifecycle involves reverse transcription of the viral genomic RNA (gRNA). Human tRNA(LYs3) serves as a primer for transcription initiation and is selectively enriched in virus particles. Human lysyl-tRNA synthetase (hLysRS) is also packaged into virions. Recently, a tRNA-like element (TLE) within the HIV-1 gRNA was shown to mimic the global tRNA fold and bind competitively to hLysRS, suggesting a mechanism of tRNA targeting to the primer binding site (PBS) and release from the synthetase. Here, we use NMR to investigate hLysRS anticodon-binding domain (ACB) binding to six RNA oligonucleotides, including a hairpin derived from the HIV-1 gRNA TLE. We show that ACB interacts with submicromolar affinity to U-rich RNA oligonucleotides the tRNALYs3 anticodon stem loop (ACSL), the WT TLE, and a nonanucleotide, U9. In contrast, the ACB bound only weakly to two TLE loop mutants and a C9 nonanucleotide. NMR chemical shift perturbations induced by each RNA indicate that the ACSL and the WT TLE both interact with the ACB in a strikingly similar manner. Taken together, these findings support the conclusion that tRNA mimicry by the HIV-1 genome leads to a highly specific protein RNA interaction that facilitates efficient primer release from hLysRS prior to reverse transcription.

• 出版日期2016-12
• 单位NIH