Characterization of association of human mitochondrial lysyl-tRNA synthetase with HIV-1 Pol and tRNA3Lys

Khoder-Agha, Fawzi; Dias, José; Comisso, Martine; Mirande, Marc

doi:10.1186/s12858-018-0092-x

Cited by 6 publications

(10 citation statements)

References 33 publications

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“…Recently, the molecular details of the interaction between mLysRS and Gag-Pol have been revealed (Phongsavanh et al, 2020). The C-terminal integrase subunit of Gag-Pol is shown to stabilize the tRNA Lys,3 :mLysRS interaction (Khoder-Agha et al, 2018;Phongsavanh et al, 2020).…”

Section: Trna Prime Reverse Transcription Of Retroviruses and Retrotrmentioning

confidence: 99%

Hijacking tRNAs From Translation: Regulatory Functions of tRNAs in Mammalian Cell Physiology

Avcilar-Kucukgoze

Kashina

2020

Front. Mol. Biosci.

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Transfer tRNAs (tRNAs) are small non-coding RNAs that are highly conserved in all kingdoms of life. Originally discovered as the molecules that deliver amino acids to the growing polypeptide chain during protein synthesis, tRNAs have been believed for a long time to play exclusive role in translation. However, recent studies have identified key roles for tRNAs and tRNA-derived small RNAs in multiple other processes, including regulation of transcription and translation, posttranslational modifications, stress response, and disease. These emerging roles suggest that tRNAs may be central players in the complex machinery of biological regulatory pathways. Here we overview these non-canonical roles of tRNA in normal physiology and disease, focusing largely on eukaryotic and mammalian systems.

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Section: Trna Prime Reverse Transcription Of Retroviruses and Retrotrmentioning

confidence: 99%

Hijacking tRNAs From Translation: Regulatory Functions of tRNAs in Mammalian Cell Physiology

Avcilar-Kucukgoze

Kashina

2020

Front. Mol. Biosci.

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“…These three constructs contain the CCD dimerization domain of IN. The apparent dissociation constants of the complexes formed between these three integrase species and mLysRS were determined using a homogeneous time-resolved fluorescence (HTRF) assay described in Khoder-Agha et al [ 12 ]. Whereas the binding affinity determined for IN-∆N-H 6 ( K d of 3.0 ± 0.4 nM) was similar to that determined for native integrase (IN-H 6 ; K d of 2.6 ± 0.3 nM), no interaction could be detected with CCD-H 6 ( K d > 400 nM) ( Figure 2 ).…”

Section: Resultsmentioning

confidence: 99%

“…The packaging of tRNA 3 Lys from the host cell within newly made HIV-1 particles involves the formation of a ternary complex comprising the GagPol viral polyprotein, mitochondrial lysyl-tRNA synthetase and tRNA Lys from the host cell. The integrase domain located at the very C-terminus of the GagPol precursor protein (Figure 1) is the main contributor to the stability of the complex between mLysRS and GagPol [12]. The cryo-EM structural analysis of the HIV-1 STC intasome revealed that integrase is made of three well-defined structural domains (Figure 1).…”

Section: The C-terminal Domain Is the Major Region Of Integrase Interacting With Mlysrsmentioning

confidence: 99%

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How HIV-1 Integrase Associates with Human Mitochondrial Lysyl-tRNA Synthetase

Phongsavanh

Al-Qatabi

Shaban

et al. 2020

Viruses

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Replication of human immunodeficiency virus type 1 (HIV-1) requires the packaging of tRNALys,3 from the host cell into the new viral particles. The GagPol viral polyprotein precursor associates with mitochondrial lysyl-tRNA synthetase (mLysRS) in a complex with tRNALys, an essential step to initiate reverse transcription in the virions. The C-terminal integrase moiety of GagPol is essential for its association with mLysRS. We show that integrases from HIV-1 and HIV-2 bind mLysRS with the same efficiency. In this work, we have undertaken to probe the three-dimensional (3D) architecture of the complex of integrase with mLysRS. We first established that the C-terminal domain (CTD) of integrase is the major interacting domain with mLysRS. Using the pBpa-photo crosslinking approach, inter-protein cross-links were observed involving amino acid residues located at the surface of the catalytic domain of mLysRS and of the CTD of integrase. In parallel, using molecular docking simulation, a single structural model of complex was found to outscore other alternative conformations. Consistent with crosslinking experiments, this structural model was further probed experimentally. Five compensatory mutations in the two partners were successfully designed which supports the validity of the model. The complex highlights that binding of integrase could stabilize the tRNALys:mLysRS interaction.

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“… 18 , 28 , 29 In the virus, LysRS has been proposed to interact with Gag and Gag-Pol. 30 − 36 Specifically, LysRS has been suggested to interact with the connection and RNase H regions in the RT domain of the Gag-Pol polyprotein. 32 However, whether LysRS affects RT maturation is unknown.…”

Section: Introductionmentioning

confidence: 99%

Effect of Lysyl–tRNA Synthetase on the Maturation of HIV-1 Reverse Transcriptase

et al. 2020

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In human immunodeficiency virus-1 (HIV-1), reverse transcriptase (RT) is encoded as a 66 kDa protein, p66, in the Gag-Pol polyprotein. This protein is proteolytically cleaved by HIV-1 protease (PR) to finally generate a mature RT that is a heterodimer, composed of a p66 subunit and a p66-derived 51 kDa subunit, p51. In our prior work, we demonstrated that tRNA Lys3 binding to p66/p66 facilitates efficient cleavage of p66 to p51 by PR. However, tRNA Lys3 is known to be recruited to the virus by forming a complex with lysyl–tRNA synthetase (LysRS). Herein, we tested whether LysRS can have an effect on RT maturation in vitro . Importantly, our data show no significant differences in RT maturation in the presence of LysRS. Furthermore, no apparent p66/66 interaction with LysRS was observed. Although PR cleaved LysRS, it did not immediately release tRNA Lys3 from LysRS. Thus, we conclude that a free fraction of tRNA Lys3 , which is in equilibrium with a LysRS-bound form, interacts with p66/p66 without any additional mechanism involving release of tRNA Lys3 from LysRS. Given that only transient tRNA Lys3 –p66/p66 interaction is needed for efficient RT maturation, a small amount of free tRNA may be sufficient for this process. These studies reveal molecular level insights into RT maturation and will be useful for the design of cellular/viral experiments to better understand the role of tRNA in HIV-1 replication.

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Characterization of association of human mitochondrial lysyl-tRNA synthetase with HIV-1 Pol and tRNA3Lys

Cited by 6 publications

References 33 publications

Hijacking tRNAs From Translation: Regulatory Functions of tRNAs in Mammalian Cell Physiology

Hijacking tRNAs From Translation: Regulatory Functions of tRNAs in Mammalian Cell Physiology

How HIV-1 Integrase Associates with Human Mitochondrial Lysyl-tRNA Synthetase

Effect of Lysyl–tRNA Synthetase on the Maturation of HIV-1 Reverse Transcriptase

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