title: Anticodon-like binding of the HIV-1 tRNA-like element to human lysyl-tRNA synthetase.
abstract: A critical step in the HIV-1 lifecycle involves reverse transcription of the viral genomic RNA (gRNA). Human tRNALys3serves 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 tRNALys3anticodon 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.
authors.affiliation: Department of Chemistry, University of Cincinnati, Cincinnati, Ohio 45221-0172, USA.
authors.forename: Sheng
authors.initials: S
authors.lastname: Liu
authors.affiliation: Department of Chemistry and Biochemistry, Center for Retrovirus Research and Center for RNA Biology, The Ohio State University, Columbus, Ohio 43210, USA.
authors.forename: Roopa
authors.initials: R
authors.lastname: Comandur
authors.affiliation: Department of Chemistry and Biochemistry, Center for Retrovirus Research and Center for RNA Biology, The Ohio State University, Columbus, Ohio 43210, USA.
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Jones
authors.affiliation: Department of Chemistry, University of Cincinnati, Cincinnati, Ohio 45221-0172, USA.
authors.forename: Pearl
authors.initials: P
authors.lastname: Tsang
authors.affiliation: Department of Chemistry and Biochemistry, Center for Retrovirus Research and Center for RNA Biology, The Ohio State University, Columbus, Ohio 43210, USA.
authors.forename: Karin
authors.initials: K
authors.lastname: Musier-Forsyth
mesh_terms: D000926:Anticodon; D005454:Fluorescence Polarization; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D008250:Lysine-tRNA Ligase / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D012343:RNA, Transfer / Q000235:genetics / Q000378:metabolism*
chemical_list: D000926:Anticodon; D012343:RNA, Transfer; D008250:Lysine-tRNA Ligase
keywords: HIV-1 genome; NMR; anticodon-binding domain; chemical shift perturbation; human lysyl-tRNA synthetase; tRNA-like element