Translation initiation of the HIV-1 mRNA

Ohlmann, Théophile; Mengardi, Chloé; López-Lastra, Marcelo

doi:10.4161/trla.29629

Cited by 3 publications

(3 citation statements)

References 163 publications

(160 reference statements)

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“…Due to the apparent relevance in driving HIV-1 Gag protein synthesis when cap-dependent translation initiation is hindered [23][24][25][26][27]30], in this study, we focused on understanding the mechanism of translation driven by the IRES that is located within the 5 0 UTR of the HIV-1 full length mRNA (HIV IRES), [25,26,28]. To date, the molecular basis of HIV-1 IRES-mediated translation initiation remains poorly understood (reviewed in [70,71]). The HIV-1 IRES is active in HeLa and COS-7 cells (Figs 1-4), although, relative to other viral IRESs, its activity is low in HeLa cells ( Fig.…”

Section: Discussionmentioning

confidence: 99%

Structural domains within the HIV‐1 mRNA and the ribosomal protein S25 influence cap‐independent translation initiation

et al. 2016

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The 5′leader of the HIV-1 genomic RNA is a multifunctional region that folds into secondary/tertiary structures that regulate multiple processes during viral replication including translation initiation. In this work we examine the internal ribosome entry site (IRES) located in the 5′leader that drives translation initiation of the viral Gag protein under conditions that hinder cap-dependent translation initiation. We show that activity of the HIV-1 IRES relies on ribosomal protein S25 (eS25). Additionally, a mechanistic and mutational analysis revealed that the HIV-1 IRES is modular in nature and that once the 40S ribosomal subunit is recruited to the IRES, translation initiates without the need of ribosome scanning. These findings elucidate a mechanism of initiation by the HIV-1 IRES whereby a number of highly structured sites present within the HIV-1 5′leader leads to the recruitment of the 40S subunit directly at the site of initiation of protein synthesis.

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Section: Discussionmentioning

confidence: 99%

Structural domains within the HIV‐1 mRNA and the ribosomal protein S25 influence cap‐independent translation initiation

et al. 2016

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“…3,5 Posisi IRES HIV-1 diketahui terletak pada 5'UTR. 6 Brasey et al melaporkan bahwa aktivitas IRES minimal terdapat pada basa ke 104-336 dari awal 5'UTR dengan 4 domain penting yaitu primer binding site (PBS), dimerization site (DIS), splice donor site (SD), dan packaging signal (Ψ). 3 Aktivitas IRES 5'UTR HIV-1d dilaporkan tinggi dalam mengekspresikan gen pada sel tropisme dari HIV-1 yaitu sel line THP-1 (monosit) dan sel line HPB-ALL (limfosit).…”

Section: Pendahuluanunclassified

Konstruksi Plasmid Rekombinan untuk Inisiasi Translasi Enhance Green Fluorescent Protein oleh Internal Ribosomal Entry Site HIV-1

Rahmagiarti

Widyaningtyas

Bela

2018

mpk

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Human immunodeficiency virus (HIV) is a virus that causes acquired immunodeficiency virus syndrome (AIDS). The HIV genome has a cap structure at 5’ and polyadenylation at 3’ on mRNA resulting in a translation initiation through scanning at 5'untranslated region (UTR). The Vpr protein produced during viral replication causes the 5'cap scanning to be inhibited so HIV-1 can directly recruit the ribosome at the start codon via internal ribosomal entry site (IRES). IRES activity is high at G2/M phase and highest expression in monocyte cell line (THP-1) and lymphocyte (HPB-ALL). The role of HIV IRES however, is not yet known in infection of nondividing cells by HIV-1. HIV-1 IRES and egfp from pcDNA5FRT/TO were amplified with PCR. The insert DNA (HIV-1 IRES_egfp) and pcDNA3.1(+) were digested with EcoRI and ApaI and then ligated. The verification was performed with PCR colonies, restriction verification, and sequencing. The size of insert DNA is 1067 bp while the vector is 5379 bp. E. coli transformed with DNA ligation produces 70 colonies, control of ligation produces 5 colonies, and negative control didn’t grow. 19 colonies contain recombinant DNA, restriction verification was of the appropriate size, and the sequence verification didn’t find any mutation. Therefore, the subcloning process pcDNA3.1_IRES HIV-1_egfp was successfully performed. Abstrak Human immunodeficiency virus (HIV) merupakan virus penyebab acquired immunodeficiency virus syndrome (AIDS). Genom HIV memiliki struktur cap di 5’ dan poliadenilasi di 3’ mRNA sehingga proses inisiasi translasi melalui pemindaian 5’cap pada struktur untranslated region (UTR) di 5’ mRNA HIV. Protein Vpr yang dihasilkan selama replikasi virus menyebabkan pemindaian melalui 5’cap terhambat sehingga HIV-1 dapat langsung merekrut ribosom pada kodon awal translasi melalui struktur internal ribosomal entry site (IRES). Aktivitas IRES tinggi pada fase G2/M dan ekspresi gen tinggi pada sel line monosit (THP-1) dan limfosit (HPB-ALL). Namun, peran IRES HIV-1 belum diketahui pada sel tidak membelah yang merupakan sel target pada infeksi HIV-1. DNA sekuen IRES HIV-1 dan egfp dari pcDNA5FRT/TO diamplifikasi dengan PCR. DNA sisipan (IRES HIV-1_egfp) dan pcDNA3.1(+) dipotong dengan EcoRI dan ApaI lalu DNA sisipan diligasi dengan pcDNA3.1(+). Verifikasi hasil klona dilakukan dengan PCR koloni, verifikasi restriksi, dan sekuensing. Restriksi DNA sisipan menghasilkan pita berukuran 1067 pb. Restriksi vektor plasmid menghasilkan pita berukuran 5379 pb. E.coli yang ditransformasi DNA ligasi menghasilkan 70 koloni, kontrol ligasi 5 koloni, dan kontrol negatif tidak tumbuh. 19 koloni terverifikasi mengandung DNA rekombinan, verifikasi restriksi memiliki ukuran sesuai, dan verifikasi sekuensing tidak terdapat perubahan basa. Oleh karena itu, proses subkloning pcDNA3.1_IRES HIV-1_egfp berhasil dilakukan.

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“…Ribosome recruitment onto the HIV-1 unspliced mRNA has been shown to occur through capdependent and cap-independent mechanisms (1)(2)(3). Initial studies showed that the highly structured 5´-untranslated region (5´-UTR) present within the full-length unspliced mRNA was inhibitory for translation in cell-free translation systems suggesting that cap-dependent ribosome scanning was not an efficient mechanism of translation initiation operating in HIV-1 transcripts (4)(5)(6).…”

Section: Introductionmentioning

confidence: 99%

CBP80/20-dependent translation initiation factor (CTIF) inhibits HIV-1 Gag synthesis by targeting the function of the viral protein Rev

et al. 2020

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Translation initiation of the human immunodeficiency virus type-1 (HIV-1) unspliced mRNA has been shown to occur through cap-dependent and IRES-driven mechanisms. Previous studies suggested that the nuclear cap-binding complex (CBC) rather than eIF4E drives cap-dependent translation of the unspliced mRNA and we have recently reported that the CBC subunit CBP80 supports the function of the viral protein Rev during nuclear export and translation of this viral transcript. Ribosome recruitment during CBC-dependent translation of cellular mRNAs relies on the activity CBP80/20 translation initiation factor (CTIF), which bridges CBP80 and the 40S ribosomal subunit through interactions with eIF3g. Here, we report that CTIF restricts HIV-1 replication by interfering with Gag synthesis from the unspliced mRNA. Our results indicate that CTIF associates with Rev through its N-terminal domain and is recruited onto the unspliced mRNA ribonucleoprotein complex in order to block translation. We also demonstrate that CTIF induces the cytoplasmic accumulation of Rev impeding the association of the viral protein with CBP80. We finally show that CTIF restricts HIV-2 but not MLV Gag synthesis indicating an inhibitory mechanism conserved in Rev-expressing human lentiviruses..

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Translation initiation of the HIV-1 mRNA

Cited by 3 publications

References 163 publications

Structural domains within the HIV‐1 mRNA and the ribosomal protein S25 influence cap‐independent translation initiation

Structural domains within the HIV‐1 mRNA and the ribosomal protein S25 influence cap‐independent translation initiation

Konstruksi Plasmid Rekombinan untuk Inisiasi Translasi Enhance Green Fluorescent Protein oleh Internal Ribosomal Entry Site HIV-1

CBP80/20-dependent translation initiation factor (CTIF) inhibits HIV-1 Gag synthesis by targeting the function of the viral protein Rev

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