Molecular Investigations of Ankyrin Specific to HIV-1 Capsid in Inhibition of Viral Replication and Direct Cell-to-Cell Viral Transfer

A designed ankyrin repeat scaffold protein (AnkGAG1D4), which recognizes the human immunodeficiency virus-1 (HIV-1) capsid (CA) protein, was formerly established and described as an HIV-1 inhibitor that interfere the assembly process of HIV-1 replication cycle resulting in the significant reduction...

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Main Author: Sutpirat Moonmuang
Other Authors: Prof. Dr. Chatchai Tayapiwatana
Format: Theses and Dissertations
Language:English
Published: เชียงใหม่ : บัณฑิตวิทยาลัย มหาวิทยาลัยเชียงใหม่ 2020
Online Access:http://cmuir.cmu.ac.th/jspui/handle/6653943832/69601
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Institution: Chiang Mai University
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spelling th-cmuir.6653943832-696012020-08-17T01:44:39Z Molecular Investigations of Ankyrin Specific to HIV-1 Capsid in Inhibition of Viral Replication and Direct Cell-to-Cell Viral Transfer การศึกษาบทบาทระดับโมเลกุลของโปรตีนแองไครินที่จำเพาะต่อไวรัสเอชไอวี-1 แคปซิดในการยับยั้งการเพิ่มจำนวนของเชื้อ และการส่งผ่านของไวรัสโดยตรงจากเซลล์สู่เซลล์ Sutpirat Moonmuang Prof. Dr. Chatchai Tayapiwatana Dr. Umpa Yasamut Dr. Supachai Sakkhachornphop A designed ankyrin repeat scaffold protein (AnkGAG1D4), which recognizes the human immunodeficiency virus-1 (HIV-1) capsid (CA) protein, was formerly established and described as an HIV-1 inhibitor that interfere the assembly process of HIV-1 replication cycle resulting in the significant reduction of viral progeny. However, the details of mode and molecular mechanisms of action of AnkGAG1D4 during the late stages of the HIV-1 replication cycle have not been investigated. Using stimulated emission-depletion microscopy, we demonstrated that Gag distribution was clearly interrupted at the inner cell membrane leaflets of HIV-1 transfected cells. This implied that Gag polymerization was disturbed by AnkGAG1D4. Disturbing of Gag polymerization resulted in the reduction of viral release without effecting proteolytic processing of Gag as determined by measuring the HIV-1 protease activity, induction of Gag accumulation inside HIV-1 producer cells, and reduction of the CD81 tetraspanin organization at the plasma membrane of HIV-1 transfected cell. Moreover, reverse transcription-quantitative polymerase chain reaction experiments were performed to determine levels of viral and cellular RNAs, and the packaging efficiency of RNAs. Our results revealed that AnkGAG1D4 impeded the Gag precursor protein from packaging of viral and cellular RNAs for encapsidation into viral particles. Finally, the intercellular transmission of HIV-1 through direct cell-to-cell contact was established. The cell-to-cell transmission of HIV-1 from Sup-T1 to THP-1 was considerably prohibited by myristoylated AnkGAG1D4. These findings convey additional information on the antiviral activity of AnkGAG1D4 at late stages of the HIV-1 replication cycle which will be valuable for either protein scaffold or chemical drug design to prevent the HIV assembly at inner membrane. 2020-08-17T01:44:39Z 2020-08-17T01:44:39Z 2020-06 Thesis http://cmuir.cmu.ac.th/jspui/handle/6653943832/69601 en เชียงใหม่ : บัณฑิตวิทยาลัย มหาวิทยาลัยเชียงใหม่
institution Chiang Mai University
building Chiang Mai University Library
continent Asia
country Thailand
Thailand
content_provider Chiang Mai University Library
collection CMU Intellectual Repository
language English
description A designed ankyrin repeat scaffold protein (AnkGAG1D4), which recognizes the human immunodeficiency virus-1 (HIV-1) capsid (CA) protein, was formerly established and described as an HIV-1 inhibitor that interfere the assembly process of HIV-1 replication cycle resulting in the significant reduction of viral progeny. However, the details of mode and molecular mechanisms of action of AnkGAG1D4 during the late stages of the HIV-1 replication cycle have not been investigated. Using stimulated emission-depletion microscopy, we demonstrated that Gag distribution was clearly interrupted at the inner cell membrane leaflets of HIV-1 transfected cells. This implied that Gag polymerization was disturbed by AnkGAG1D4. Disturbing of Gag polymerization resulted in the reduction of viral release without effecting proteolytic processing of Gag as determined by measuring the HIV-1 protease activity, induction of Gag accumulation inside HIV-1 producer cells, and reduction of the CD81 tetraspanin organization at the plasma membrane of HIV-1 transfected cell. Moreover, reverse transcription-quantitative polymerase chain reaction experiments were performed to determine levels of viral and cellular RNAs, and the packaging efficiency of RNAs. Our results revealed that AnkGAG1D4 impeded the Gag precursor protein from packaging of viral and cellular RNAs for encapsidation into viral particles. Finally, the intercellular transmission of HIV-1 through direct cell-to-cell contact was established. The cell-to-cell transmission of HIV-1 from Sup-T1 to THP-1 was considerably prohibited by myristoylated AnkGAG1D4. These findings convey additional information on the antiviral activity of AnkGAG1D4 at late stages of the HIV-1 replication cycle which will be valuable for either protein scaffold or chemical drug design to prevent the HIV assembly at inner membrane.
author2 Prof. Dr. Chatchai Tayapiwatana
author_facet Prof. Dr. Chatchai Tayapiwatana
Sutpirat Moonmuang
format Theses and Dissertations
author Sutpirat Moonmuang
spellingShingle Sutpirat Moonmuang
Molecular Investigations of Ankyrin Specific to HIV-1 Capsid in Inhibition of Viral Replication and Direct Cell-to-Cell Viral Transfer
author_sort Sutpirat Moonmuang
title Molecular Investigations of Ankyrin Specific to HIV-1 Capsid in Inhibition of Viral Replication and Direct Cell-to-Cell Viral Transfer
title_short Molecular Investigations of Ankyrin Specific to HIV-1 Capsid in Inhibition of Viral Replication and Direct Cell-to-Cell Viral Transfer
title_full Molecular Investigations of Ankyrin Specific to HIV-1 Capsid in Inhibition of Viral Replication and Direct Cell-to-Cell Viral Transfer
title_fullStr Molecular Investigations of Ankyrin Specific to HIV-1 Capsid in Inhibition of Viral Replication and Direct Cell-to-Cell Viral Transfer
title_full_unstemmed Molecular Investigations of Ankyrin Specific to HIV-1 Capsid in Inhibition of Viral Replication and Direct Cell-to-Cell Viral Transfer
title_sort molecular investigations of ankyrin specific to hiv-1 capsid in inhibition of viral replication and direct cell-to-cell viral transfer
publisher เชียงใหม่ : บัณฑิตวิทยาลัย มหาวิทยาลัยเชียงใหม่
publishDate 2020
url http://cmuir.cmu.ac.th/jspui/handle/6653943832/69601
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