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USP49 potently stabilizes APOBEC3G protein by removing ubiquitin and inhibits HIV-1 replication.

Ting Pan | Zheng Song | Liyang Wu | Guangyan Liu | Xiancai Ma | Zhilin Peng | Mo Zhou | Liting Liang | Bingfeng Liu | Jun Liu | Junsong Zhang | Xuanhong Zhang | Ryan Huang | Jiacong Zhao | Yonghong Li | Xuemei Ling | Yuewen Luo | Xiaoping Tang | Weiping Cai | Kai Deng | Linghua Li | Hui Zhang
eLife | 2019

The antiviral activity of host factor apolipoprotein B mRNA editing enzyme catalytic polypeptide-like 3G (APOBEC3G, A3G) and its degradation mediated by human immunodeficiency virus type 1 (HIV-1) Vif protein are important topics. Although accumulating evidence indicates the importance of deubiquitination enzymes (DUBs) in innate immunity, it is unknown if they participate in A3G stability. Here, we found that USP49 directly interacts with A3G and efficiently removes ubiquitin, consequently increasing A3G protein expression and significantly enhancing its anti-HIV-1 activity. Unexpectedly, A3G degradation was also mediated by a Vif- and cullin-ring-independent pathway, which was effectively counteracted by USP49. Furthermore, clinical data suggested that USP49 is correlated with A3G protein expression and hypermutations in Vif-positive proviruses, and inversely with the intact provirus ratio in the HIV-1 latent reservoir. Our studies demonstrated a mechanism to effectively stabilize A3G expression, which could comprise a target to control HIV-1 infection and eradicate the latent reservoir.

Pubmed ID: 31397674

Associated grants

  • Agency: National Special Research Program of China for Important Infectious Diseases, International
    Id: 2018ZX10302103
  • Agency: National Special Research Program of China for Important Infectious Diseases, International
    Id: 2017ZX10202102
  • Agency: National Natural Science Foundation of China, International
    Id: Key Program 81730060
  • Agency: The International Collaboration Program of the Natural Science Foundation of China and the US NIH, International
    Id: 81561128007
  • Agency: The Joint-Innovation Program in Healthcare for Special Scientific Research Projects of Guangzhou, International
    Id: 201803040002
  • Agency: The Science and Technology Planning Project of Guangzhou, International
    Id: 201704020226
  • Agency: Pearl River S and T Nova Program of Guangzhou, International
    Id: 201806010118
  • Agency: National Natural Science Foundation of China, International
    Id: 81971918
  • Agency: National Special Research Program of China for Important Infectious Diseases, International
    Id: 2018ZX10302103; 2017ZX10202102
  • Agency: The Important Key Program of the Natural Science Foundation of China, International
    Id: 81730060
  • Agency: The International Collaboration Program of the Natural Science Foundation of China and the U.S. NIH, International
    Id: 81561128007
  • Agency: National Natural Science Foundation of China, International
    Id: 8197080527

Publication data is provided by the National Library of Medicine ® and PubMed ®. Data is retrieved from PubMed ® on a weekly schedule. For terms and conditions see the National Library of Medicine Terms and Conditions.

This is a list of tools and resources that we have found mentioned in this publication.


ATCC (tool)

RRID:SCR_001672

Global nonprofit biological resource center (BRC) and research organization that provides biological products, technical services and educational programs to private industry, government and academic organizations. Its mission is to acquire, authenticate, preserve, develop and distribute biological materials, information, technology, intellectual property and standards for the advancement and application of scientific knowledge. The primary purpose of ATCC is to use its resources and experience as a BRC to become the world leader in standard biological reference materials management, intellectual property resource management and translational research as applied to biomaterial development, standardization and certification. ATCC characterizes cell lines, bacteria, viruses, fungi and protozoa, as well as develops and evaluates assays and techniques for validating research resources and preserving and distributing biological materials to the public and private sector research communities.

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Addgene (tool)

RRID:SCR_002037

Non-profit plasmid repository dedicated to helping scientists around the world share high-quality plasmids. Facilitates archiving and distributing DNA-based research reagents and associated data to scientists worldwide. Repository contains over 65,000 plasmids, including special collections on CRISPR, fluorescent proteins, and ready-to-use viral preparations. There is no cost for scientists to deposit plasmids, which saves time and money associated with shipping plasmids themselves. All plasmids are fully sequenced for validation and sequencing data is openly available. We handle the appropriate Material Transfer Agreements (MTA) with institutions, facilitating open exchange and offering intellectual property and liability protection for depositing scientists. Furthermore, we curate free educational resources for the scientific community including a blog, eBooks, video protocols, and detailed molecular biology resources.

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Ensembl (tool)

RRID:SCR_002344

Collection of genome databases for vertebrates and other eukaryotic species with DNA and protein sequence search capabilities. Used to automatically annotate genome, integrate this annotation with other available biological data and make data publicly available via web. Ensembl tools include BLAST, BLAT, BioMart and the Variant Effect Predictor (VEP) for all supported species.

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PRISM (tool)

RRID:SCR_005375

THIS RESOURCE IS NO LONGER IN SERVICE. Documented on May 5,2022.Tool that predicts interactions between transcription factors and their regulated genes from binding motifs. Understanding vertebrate development requires unraveling the cis-regulatory architecture of gene regulation. PRISM provides accurate genome-wide computational predictions of transcription factor binding sites for the human and mouse genomes, and integrates the predictions with GREAT to provide functional biological context. Together, accurate computational binding site prediction and GREAT produce for each transcription factor: 1. putative binding sites, 2. putative target genes, 3. putative biological roles of the transcription factor, and 4. putative cis-regulatory elements through which the factor regulates each target in each functional role.

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BD Biosciences (tool)

RRID:SCR_013311

An Antibody supplier

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Quantity One 1-D Analysis Software (tool)

RRID:SCR_014280

Software used for Bio-Rad imaging systems to acquire, quantitate, and analyze a variety of data. The software allows automatic configuration of imaging systems with appropriate filters, lasers, LEDs, and other illumination sources. It also contains tools for automated analysis of tests and assays such as 1-D electrophoretic gels, western blots, and colony counts.

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Odyssey CLx (tool)

RRID:SCR_014579

A data analysis software developed to simplify and record data from fluorescent related assays such as Western Blots, EMSAs, and Western Assays. This software will consolidate all of the resulting data into one easy-to-track image, and thus removes the need for multiple exposures, as well as standardizing the resulting fluorescent bands without image saturation, blowout, or removed sensitivity. Odyssey CLx is capable of scanning multiple mini-blots, microplates, or slides at the same time.

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NIH AIDS Reagent Program (tool)

RRID:SCR_023191

NIH HIV Reagent Program has been managed under contract by American Type Culture Collection (ATCC) since 2020. ATCC shall maintain the NIH HIV Reagent Program through identification, acquisition, production, receipt, storage, maintenance, distribution and disposal of biological and chemical research organisms and materials for HIV and other infectious diseases for use in basic and translational research.

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U-118MG (tool)

RRID:CVCL_0633

Cell line U-118MG is a Cancer cell line with a species of origin Homo sapiens (Human)

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RRID:AB_591224

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Beta Actin antibody (antibody)

RRID:AB_2687938

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Goat anti-Mouse IgG H&L (DyLight® 594) secondary antibody (antibody)

RRID:AB_10680252

This polyclonal targets Goat anti-Mouse IgG H&L (DyLight® 594) secondary antibody

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Goat polyclonal Secondary Antibody to Mouse IgG - H&L (DyLight® 488), pre-adsorbed (antibody)

RRID:AB_10687475

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USP18 (D4E7) Rabbit mAb (antibody)

RRID:AB_10614342

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RRID:AB_2263076

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ubiquitin antibody (antibody)

RRID:AB_671515

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RRID:AB_10063035

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EZview(TM) Red Anti-HA Affinity Gel (antibody)

RRID:AB_10109562

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USP49 antibody (antibody)

RRID:AB_10643992

This polyclonal targets USP49

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anti-HA-tag (antibody)

RRID:AB_10951811

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APOBEC3G antibody [mAbcam75560] (antibody)

RRID:AB_10676129

This monoclonal targets APOBEC3G antibody [mAbcam75560]

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IRDye 800CW Goat anti-Rabbit IgG (antibody)

RRID:AB_621843

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GraphPad Prism (software resource)

RRID:SCR_002798

Statistical analysis software that combines scientific graphing, comprehensive curve fitting (nonlinear regression), understandable statistics, and data organization. Designed for biological research applications in pharmacology, physiology, and other biological fields for data analysis, hypothesis testing, and modeling.

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Image Studio Lite (software resource)

RRID:SCR_013715

THIS RESOURCE IS NO LONGER IN SERVICE. Documented on September 28,2023. Software application which can analyze basic Western blot data and export them for use in spreadsheet programs. Users can create standardized or custom lab reports and can share them with colleagues. Image Studio Lite has been discontinued and replaced with Empiria Studio Software.

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Hypermut (software resource)

RRID:SCR_014933

Web application for the analysis and detection of APOBEC-induced hypermutations. The first sequence in the input alignment will be used as the reference sequence, and each of the other sequences will be used as a query sequence.

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FlowJo (software resource)

RRID:SCR_008520

Software for single-cell flow cytometry analysis. Its functions include management, display, manipulation, analysis and publication of the data stream produced by flow and mass cytometers.

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HeLa (cell line)

RRID:CVCL_0030

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HEK293T (cell line)

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