Cooperative methylation of human tRNA3Lys at positions A58 and U54 drives the early and late steps of HIV-1 replication. Issue 20 (13th October 2021)

• Record Type: Journal Article
• Authors: Fukuda, Hiroyuki; Chujo, Takeshi; Wei, Fan-Yan; Shi, Sheng-Lan; Hirayama, Mayumi; Kaitsuka, Taku; Yamamoto, Takahiro; Oshiumi, Hiroyuki; Tomizawa, Kazuhito
• Is Part Of: Nucleic acids research. Volume 49:Issue 20(2021)
• Page Start: 11855
• Page End: 11867
• Publication Date: 2021-10-13
• Subjects: Nucleic acids -- Periodicals; Molecular biology -- Periodicals; 572.805
• DOI: 10.1093/nar/gkab879
• Languages: English
• ISSNs: 0305-1048

Abstract

Abstract: Retroviral infection requires reverse transcription, and the reverse transcriptase (RT) uses cellular tRNA as its primer. In humans, the TRMT6-TRMT61A methyltransferase complex incorporates N 1 -methyladenosine modification at tRNA position 58 (m 1 A58); however, the role of m 1 A58 as an RT-stop site during retroviral infection has remained questionable. Here, we constructed TRMT6 mutant cells to determine the roles of m 1 A in HIV-1 infection. We confirmed that tRNA3 Lys m 1 A58 was required for in vitro plus-strand strong-stop by RT. Accordingly, infectivity of VSV-G pseudotyped HIV-1 decreased when the virus contained m 1 A58-deficient tRNA3 Lys instead of m 1 A58-modified tRNA3 Lys . In TRMT6 mutant cells, the global protein synthesis rate was equivalent to that of wild-type cells. However, unexpectedly, plasmid-derived HIV-1 expression showed that TRMT6 mutant cells decreased accumulation of HIV-1 capsid, integrase, Tat, Gag, and GagPol proteins without reduction of HIV-1 RNAs in cells, and fewer viruses were produced. Moreover, the importance of 5, 2′- O -dimethyluridine at U54 of tRNA3 Lys as a second RT-stop site was supported by conservation of retroviral genome-tRNA Lys sequence-complementarity, and TRMT6 was required for efficient 5-methylation of U54. These findings illuminate the fundamental importance of tRNA m 1 A58 modification in both the early and late steps of HIV-1 replication, as well as in the cellular tRNA modification network.