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RNA Interference, Understanding the Collateral Effects of HIV-1 and ZIKV.

Record Type:	Electronic resources : Monograph/item
Title/Author:	RNA Interference, Understanding the Collateral Effects of HIV-1 and ZIKV./
Author:	Alpuche Lazcano, Sergio Paulo.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2019,
Description:	279 p.
Notes:	Source: Dissertations Abstracts International, Volume: 82-10, Section: B.
Contained By:	Dissertations Abstracts International82-10B.
Subject:	MicroRNAs. -
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=28266825
ISBN:	9798708719836

RNA Interference, Understanding the Collateral Effects of HIV-1 and ZIKV.
Alpuche Lazcano, Sergio Paulo.

RNA Interference, Understanding the Collateral Effects of HIV-1 and ZIKV. - Ann Arbor : ProQuest Dissertations & Theses, 2019 - 279 p.

Source: Dissertations Abstracts International, Volume: 82-10, Section: B.

Thesis (Ph.D.)--McGill University (Canada), 2019.

This item must not be sold to any third party vendors.

Eukaryotic cells possess different mechanisms of gene regulation at various stages from transcription to translation. RNA interference (RNAi) is a mechanism of post-transcriptional gene modulation based on short double-stranded RNA sequences called micro RNAs (miRNA). MiRNAs are partially complementary to a specific sequence in messenger RNAs (mRNA) and their binding to a targeted mRNA mediates its repression and/or degradation. In humans, RNAi controls over 50 % of protein-coding genes. The improper functioning of this process leads to the development of different pathologies. Internal or external factors influence the malfunctioning of RNAi. Among them, viral infections fine-tune RNAi, which can be beneficial or detrimental for viral replication.Human immunodeficiency virus type-1 (HIV-1) and Zika virus (ZIKV) are two different viruses that have distinct mechanisms of infection. Nonetheless, both viruses change the miRNA landscape through modifications in the RNAi pathway. This thesis explores the interplay between RNAi and either HIV-1 or ZIKV. In the first project, we re-evaluated the relationship between RNAi and HIV-1. Our experiments demonstrated that RNAi is functional in HIV-1 replicating cells. Besides, proteins of the RNA induced silencing complex (RISC) are not relocalized in HIV-1 producing cells. In contrast, we showed that HIV-1 Gag interacts specifically with Dicer, an essential protein of the RISC. We found that Gag does not change the cleavage of precursor. miRNAs by Dicer but modifies their availability. Gag modifies the loading of specific miRNAs on Dicer, which leads to sequestration or higher loading of particular miRNAs. Among these miRNAs, we found miR-642a 3p that has been predicted to target the AFF4 protein. AFF4 is an essential component of the virus transcription and its absence profoundly impairs HIV-1 replication.Our second goal was to examine mRNAs and miRNAs in an integrative analysis of infected cells with ZIKV. Because little information on the neurological consequences after ZIKV infection were available at the beginning of our study, we first compared two different viral strains. We determined that a Brazilian isolate was more cytopathic than an early Asian one in distinct cell types. Then, using the Brazilian virus, we studied the impact of ZIKV infection on the expression of host mRNAs and miRNAs in fetal mice neurons. Notably, we determined a downregulation of NPAS4 and NR4A transcripts, which have been linked to the neurogenesis process. Moreover, our integrative analysis showed a correlation between dysregulated mRNAs and miRNAs in the infected cells. Particularly, NR4A3 transcripts might be controlled by the dysregulated levels of miR-7116-5p and miR-7013-5p upon the infection.The results presented in this thesis contribute to the understanding of 1) HIV-1-induced modification of miRNAs loaded on Dicer and their consequences on mRNAs expression and 2) ZIKV cytopathicity and its impact on miRNAs and mRNAs expression profile in murine fetal neurons.

ISBN: 9798708719836Subjects--Topical Terms:

2147037
MicroRNAs.
Subjects--Index Terms:

RNA interference

RNA Interference, Understanding the Collateral Effects of HIV-1 and ZIKV.
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245 1 0 $a RNA Interference, Understanding the Collateral Effects of HIV-1 and ZIKV.
260 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 2019
300 $a 279 p.
500 $a Source: Dissertations Abstracts International, Volume: 82-10, Section: B.
500 $a Advisor: Gatignol, Anne;Mouland, Andrew J.
502 $a Thesis (Ph.D.)--McGill University (Canada), 2019.
506 $a This item must not be sold to any third party vendors.
520 $a Eukaryotic cells possess different mechanisms of gene regulation at various stages from transcription to translation. RNA interference (RNAi) is a mechanism of post-transcriptional gene modulation based on short double-stranded RNA sequences called micro RNAs (miRNA). MiRNAs are partially complementary to a specific sequence in messenger RNAs (mRNA) and their binding to a targeted mRNA mediates its repression and/or degradation. In humans, RNAi controls over 50 % of protein-coding genes. The improper functioning of this process leads to the development of different pathologies. Internal or external factors influence the malfunctioning of RNAi. Among them, viral infections fine-tune RNAi, which can be beneficial or detrimental for viral replication.Human immunodeficiency virus type-1 (HIV-1) and Zika virus (ZIKV) are two different viruses that have distinct mechanisms of infection. Nonetheless, both viruses change the miRNA landscape through modifications in the RNAi pathway. This thesis explores the interplay between RNAi and either HIV-1 or ZIKV. In the first project, we re-evaluated the relationship between RNAi and HIV-1. Our experiments demonstrated that RNAi is functional in HIV-1 replicating cells. Besides, proteins of the RNA induced silencing complex (RISC) are not relocalized in HIV-1 producing cells. In contrast, we showed that HIV-1 Gag interacts specifically with Dicer, an essential protein of the RISC. We found that Gag does not change the cleavage of precursor. miRNAs by Dicer but modifies their availability. Gag modifies the loading of specific miRNAs on Dicer, which leads to sequestration or higher loading of particular miRNAs. Among these miRNAs, we found miR-642a 3p that has been predicted to target the AFF4 protein. AFF4 is an essential component of the virus transcription and its absence profoundly impairs HIV-1 replication.Our second goal was to examine mRNAs and miRNAs in an integrative analysis of infected cells with ZIKV. Because little information on the neurological consequences after ZIKV infection were available at the beginning of our study, we first compared two different viral strains. We determined that a Brazilian isolate was more cytopathic than an early Asian one in distinct cell types. Then, using the Brazilian virus, we studied the impact of ZIKV infection on the expression of host mRNAs and miRNAs in fetal mice neurons. Notably, we determined a downregulation of NPAS4 and NR4A transcripts, which have been linked to the neurogenesis process. Moreover, our integrative analysis showed a correlation between dysregulated mRNAs and miRNAs in the infected cells. Particularly, NR4A3 transcripts might be controlled by the dysregulated levels of miR-7116-5p and miR-7013-5p upon the infection.The results presented in this thesis contribute to the understanding of 1) HIV-1-induced modification of miRNAs loaded on Dicer and their consequences on mRNAs expression and 2) ZIKV cytopathicity and its impact on miRNAs and mRNAs expression profile in murine fetal neurons.
520 $a Les cellules eucaryotes possedent differents mecanismes de regulation genique a la fois au niveau transcriptionnel et traductionnel. L'interference ARN (iARN) est un mecanisme de modulation de l'expression post-transcriptionnelle des genes impliquant de petits ARN double brin, appeles micro-ARN (miARN). Les miARN ont une complementarite imparfaite avec une sequence cible presente sur les ARN messagers (ARNm). La liaison des miARN a un ARNm induit la repression traductionnelle de ce dernier et/ou sa degradation. Chez l'humain, l'iARN controle l'expression de plus de 50% des genes codant pour des proteines. Le disfonctionnement de l'interference ARN conduit au developpement de differentes pathologies. En effet, des evenements d'origine cellulaire ou extracellulaire tels que les infections virales peuvent dereguler l'iARN, que ce soit benefique ou nefaste pour la replication virale.Le virus de l'immunodeficience humaine de type 1 (VIH-1) et le virus Zika (ZIKV) ont des mecanismes d'infection differents. Toutefois, ces deux virus modifient la composition des miARN cellulaires en perturbant la voie de l'iARN. Cette these a permis d'explorer l'interaction entre la voie de l'interference ARN et le VIH-1 ou le ZIKV. Dans le premier projet, nous avons etudie la relation entre l'iARN et le VIH-1. Nos experiences ont demontre que la voie de l'iARN reste fonctionnelle dans les cellules produisant du VIH-1. De plus, la localisation cellulaire des proteines du complexe RISC ("RNA-Induced Silencing Complex") n'est pas modifiee dans les cellules productrices du VIH-1. En revanche, nous avons montre que la proteine Gag du VIH-1 interagit avec Dicer, une proteine essentielle du complexe RISC. Nous avons constate que cette interaction proteique ne modifie pas le clivage des precurseurs de miARN mais modifie leur disponibilite. En effet, Gag modifie le chargement de miARN specifiques sur Dicer. Ceci conduit soit a une sequestration soit a une quantite plus elevee de certains miARN charges sur Dicer. Parmi ces miARN, nous avons trouve le miR-642a 3p qui ciblerait la proteine AFF4. AFF4 est une proteine essentielle a la transcription du virus et son absence altere profondement la replication du VIH-1.Notre deuxieme objectif etait d'examiner le profil d'expression des ARNm et des miARN de cellules infectees par le ZIKV. Tres peu d'informations sur les consequences neurologiques de l'infection par le ZIKV etaient disponibles au debut de notre etude. Nous avons commence par comparer deux souches virales distinctes. Ainsi, nous avons determine qu'un isolat bresilien etait plus cytopathique qu'une souche de lignee asiatique precoce et ceci dans differents types cellulaires. Par la suite, en utilisant le virus bresilien, nous avons etudie l'impact de l'infection du ZIKV sur l'expression des ARNm et des miARN cellulaires dans des neurones foetaux de souris. Nous avons determine une regulation negative des transcrits NPAS4 et NR4A, qui sont impliques dans le processus de neurogenese. De plus, notre analyse integrative a montre une correlation entre les ARNm et les miARN deregules dans les cellules infectees. En effet, lors de l'infection par le ZIKV, les transcrits de NR4A3 pourraient etre controles par les miARN miR-7116-5p et miR-7013-5p dont l'expression est deregulee.Les resultats presentes dans cette these contribuent a la comprehension:1) des modifications induites par le VIH-1 dans le chargement de miARN sur la proteine Dicer et de l'impact de ceux-ci sur l'expression des ARNm cibles et 2) de l'effet cytopathique du ZIKV et de l'impact de ce dernier sur le profil d'expression des miARN et des ARNm cellulaires dans les neurones foetaux murins.
590 $a School code: 0781.
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650 4 $a Deoxyribonucleic acid--DNA. $3 3546879
650 4 $a CRISPR. $3 3561750
650 4 $a Mammals. $3 531989
650 4 $a Human immunodeficiency virus--HIV. $3 3564909
650 4 $a Genomes. $3 592593
650 4 $a Zika virus. $3 3319806
650 4 $a Kinases. $3 3558077
650 4 $a Proteins. $3 558769
653 $a RNA interference
653 $a HIV-1
653 $a ZIKV
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710 2 $a McGill University (Canada). $3 1018122
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