Molecular Determinants in tRNA D-arm Required for Inhibition of HIV-1 Gag Membrane Binding.

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Abstrakt

Tytuł:: Molecular Determinants in tRNA D-arm Required for Inhibition of HIV-1 Gag Membrane Binding.
Autorzy:: Sumner C; Dept. of Microbiology and Immunology, University of Michigan, Ann Arbor, MI, United States.
Kotani O; Center for Pathogen Genomics, National Institute of Infectious Diseases, Tokyo, Japan.
Liu S; Dept. of Chemistry and Biochemistry, Center for Retrovirus Research, and Center for RNA Biology, The Ohio State University, Columbus, OH, United States.
Musier-Forsyth K; Dept. of Chemistry and Biochemistry, Center for Retrovirus Research, and Center for RNA Biology, The Ohio State University, Columbus, OH, United States.
Sato H; Center for Pathogen Genomics, National Institute of Infectious Diseases, Tokyo, Japan.
Ono A; Dept. of Microbiology and Immunology, University of Michigan, Ann Arbor, MI, United States. Electronic address: .
Źródło:: Journal of molecular biology [J Mol Biol] 2022 Jan 30; Vol. 434 (2), pp. 167390. Date of Electronic Publication: 2021 Dec 06.
Typ publikacji:: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't
Język:: English
Imprint Name(s):: Publication: Amsterdam : Elsevier
Original Publication: 1959- : London : Academic Press
MeSH Terms:: HIV-1/*metabolism
RNA, Transfer/*chemistry
RNA, Transfer/*metabolism
gag Gene Products, Human Immunodeficiency Virus/*chemistry
gag Gene Products, Human Immunodeficiency Virus/*metabolism
Anticodon/metabolism ; Binding Sites ; Cell Membrane/metabolism ; HIV-1/genetics ; Humans ; Molecular Docking Simulation ; Phosphatidylinositol 4,5-Diphosphate ; Protein Interaction Domains and Motifs ; RNA, Viral/genetics ; Virus Assembly/physiology
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Grant Information:: F31 AI134287 United States AI NIAID NIH HHS; R01 AI153216 United States AI NIAID NIH HHS; R37 AI071727 United States AI NIAID NIH HHS; T32 AI007528 United States AI NIAID NIH HHS
Contributed Indexing:: Keywords: acidic phospholipid; highly basic region; lipid-protein interaction; tRNA–protein interaction; virus assembly
Substance Nomenclature:: 0 (Anticodon)
0 (Phosphatidylinositol 4,5-Diphosphate)
0 (RNA, Viral)
0 (gag Gene Products, Human Immunodeficiency Virus)
9014-25-9 (RNA, Transfer)
Entry Date(s):: Date Created: 20211209 Date Completed: 20220204 Latest Revision: 20230131
Update Code:: 20240105
PubMed Central ID:: PMC8752508
DOI:: 10.1016/j.jmb.2021.167390
PMID:: 34883117
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Plasma-membrane-specific localization of Gag, an essential step in HIV-1 particle assembly, is regulated by the interaction of the Gag MA domain with PI(4,5)P 2 and tRNA-mediated inhibition of non-specific or premature membrane binding. Different tRNAs inhibit PI(4,5)P 2 -independent membrane binding to varying degrees in vitro; however, the structural determinants for this difference remain unknown. Here we demonstrate that membrane binding of full-length Gag synthesized in vitro using reticulocyte lysates is inhibited when RNAs that contain the anticodon arm of tRNA Pro , but not that of tRNA Lys3 , are added exogenously. In contrast, in the context of a liposome binding assay in which the effects of tRNAs on purified MA were tested, full-length tRNA Lys3 showed greater inhibition of MA membrane binding than full-length tRNA Pro . While transplantation of the D loop sequence of tRNA Lys3 into tRNA Pro resulted in a modest increase in the inhibitory effect relative to WT tRNA Pro , replacing the entire D arm sequence with that of tRNA Lys3 was necessary to confer the full inhibitory effects upon tRNA Pro . Together, these results demonstrate that the D arm of tRNA Lys3 is a major determinant of strong inhibition of MA membrane binding and that this inhibitory effect requires not only the D loop, which was recently reported to contact the MA highly basic region, but the loop sequence in the context of the D arm structure.
Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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