BIOMAT Database Logo BIOMAT Database Logo

Specific Interaction between eEF1A and HIV RT Is Critical for HIV-1 Reverse Transcription and a Potential Anti-HIV Target. 2015

Dongsheng Li, and Ting Wei, and Daniel J Rawle, and Fangyun Qin, and Rui Wang, and Dinesh C Soares, and Hongping Jin, and Haran Sivakumaran, and Min-Hsuan Lin, and Kirsten Spann, and Catherine M Abbott, and David Harrich
Department of Cell and Molecular Biology, QIMR Berghofer Medical Research Institute, Herston, Queensland, Australia.

Reverse transcription is the central defining feature of HIV-1 replication. We previously reported that the cellular eukaryotic elongation factor 1 (eEF1) complex associates with the HIV-1 reverse transcription complex (RTC) and the association is important for late steps of reverse transcription. Here we show that association between the eEF1 and RTC complexes occurs by a strong and direct interaction between the subunit eEF1A and reverse transcriptase (RT). Using biolayer interferometry and co-immunoprecipitation (co-IP) assays, we show that association between the eEF1 and RTC complexes occurs by a strong (KD ~3-4 nM) and direct interaction between eEF1A and reverse transcriptase (RT). Biolayer interferometry analysis of cell lysates with titrated levels of eEF1A indicates it is a predominant cellular RT binding protein. Both the RT thumb and connection domains are required for interaction with eEF1A. A single amino acid mutation, W252A, within the thumb domain impaired co-IP between eEF1A and RT, and also significantly reduced the efficiency of late reverse transcription and virus replication when incorporated into infectious HIV-1. Molecular modeling analysis indicated that interaction between W252 and L303 are important for RT structure, and their mutation to alanine did not impair heterodimerisation, but negatively impacted interaction with eEF1A. Didemnin B, which specifically binds eEF1A, potently inhibited HIV-1 reverse transcription by greater than 2 logs at subnanomolar concentrations, especially affecting reverse transcription late DNA synthesis. Analysis showed reduced levels of RTCs from HIV-1-infected HEK293T treated with didemnin B compared to untreated cells. Interestingly, HIV-1 with a W252A RT mutation was resistant to didemnin B negative effects showing that didemnin B affects HIV-1 by targeting the RT-eEF1A interaction. The combined evidence indicates a direct interaction between eEF1A and RT is crucial for HIV reverse transcription and replication, and the RT-eEF1A interaction is a potential drug target.

UI MeSH Term Description Entries
D004797 Enzyme-Linked Immunosorbent Assay An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed. ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D014779 Virus Replication The process of intracellular viral multiplication, consisting of the synthesis of PROTEINS; NUCLEIC ACIDS; and sometimes LIPIDS, and their assembly into a new infectious particle. Viral Replication,Replication, Viral,Replication, Virus,Replications, Viral,Replications, Virus,Viral Replications,Virus Replications
D015497 HIV-1 The type species of LENTIVIRUS and the etiologic agent of AIDS. It is characterized by its cytopathic effect and affinity for the T4-lymphocyte. Human immunodeficiency virus 1,HIV-I,Human Immunodeficiency Virus Type 1,Immunodeficiency Virus Type 1, Human
D015658 HIV Infections Includes the spectrum of human immunodeficiency virus infections that range from asymptomatic seropositivity, thru AIDS-related complex (ARC), to acquired immunodeficiency syndrome (AIDS). HTLV-III Infections,HTLV-III-LAV Infections,T-Lymphotropic Virus Type III Infections, Human,HIV Coinfection,Coinfection, HIV,Coinfections, HIV,HIV Coinfections,HIV Infection,HTLV III Infections,HTLV III LAV Infections,HTLV-III Infection,HTLV-III-LAV Infection,Infection, HIV,Infection, HTLV-III,Infection, HTLV-III-LAV,Infections, HIV,Infections, HTLV-III,Infections, HTLV-III-LAV,T Lymphotropic Virus Type III Infections, Human
D047468 Immunoprecipitation The aggregation of soluble ANTIGENS with ANTIBODIES, alone or with antibody binding factors such as ANTI-ANTIBODIES or STAPHYLOCOCCAL PROTEIN A, into complexes large enough to fall out of solution. Co-Immunoprecipitation,Immune Precipitation,Co Immunoprecipitation,Co-Immunoprecipitations,Immune Precipitations,Precipitation, Immune,Precipitations, Immune
D048348 Reverse Transcription The biosynthesis of DNA carried out on a template of RNA. Transcription, Reverse
D054303 HIV Reverse Transcriptase A reverse transcriptase encoded by the POL GENE of HIV. It is a heterodimer of 66 kDa and 51 kDa subunits that are derived from a common precursor protein. The heterodimer also includes an RNAse H activity (RIBONUCLEASE H, HUMAN IMMUNODEFICIENCY VIRUS) that plays an essential role the viral replication process. Reverse Transcriptase, HIV,Reverse Transcriptase, Human Immunodeficiency Virus,Transcriptase, HIV Reverse
D057809 HEK293 Cells A cell line generated from human embryonic kidney cells that were transformed with human adenovirus type 5. 293T Cells,HEK 293 Cell Line,HEK 293 Cells,Human Embryonic Kidney Cell Line 293,Human Kidney Cell Line 293,293 Cell, HEK,293 Cells, HEK,293T Cell,Cell, 293T,Cell, HEK 293,Cell, HEK293,Cells, 293T,Cells, HEK 293,Cells, HEK293,HEK 293 Cell,HEK293 Cell
D020648 Peptide Elongation Factor 1 Peptide elongation factor 1 is a multisubunit protein that is responsible for the GTP-dependent binding of aminoacyl-tRNAs to eukaryotic ribosomes. The alpha subunit (EF-1alpha) binds aminoacyl-tRNA and transfers it to the ribosome in a process linked to GTP hydrolysis. The beta and delta subunits (EF-1beta, EF-1delta) are involved in exchanging GDP for GTP. The gamma subunit (EF-1gamma) is a structural component. EF-1,Elongation Factor 1,Elongation Factor 1alpha,Elongation Factor 1beta,Elongation Factor 1delta,Elongation Factor 1gamma,EF 1,EF-1H,EF-1alpha,EF-1beta,EF-1delta,EF-1gamma,EF 1H,EF 1alpha,EF 1beta,EF 1delta,EF 1gamma

Related Publications

Dongsheng Li, and Ting Wei, and Daniel J Rawle, and Fangyun Qin, and Rui Wang, and Dinesh C Soares, and Hongping Jin, and Haran Sivakumaran, and Min-Hsuan Lin, and Kirsten Spann, and Catherine M Abbott, and David Harrich
The HIV-1 reverse transcription (RT) process as target for RT inhibitors.
March 2000, Medicinal research reviews,
Dongsheng Li, and Ting Wei, and Daniel J Rawle, and Fangyun Qin, and Rui Wang, and Dinesh C Soares, and Hongping Jin, and Haran Sivakumaran, and Min-Hsuan Lin, and Kirsten Spann, and Catherine M Abbott, and David Harrich
eEF1A demonstrates paralog specific effects on HIV-1 reverse transcription efficiency.
April 2019, Virology,
Dongsheng Li, and Ting Wei, and Daniel J Rawle, and Fangyun Qin, and Rui Wang, and Dinesh C Soares, and Hongping Jin, and Haran Sivakumaran, and Min-Hsuan Lin, and Kirsten Spann, and Catherine M Abbott, and David Harrich
HIV-1 reverse transcription initiation: a potential target for novel antivirals?
June 2008, Virus research,
Dongsheng Li, and Ting Wei, and Daniel J Rawle, and Fangyun Qin, and Rui Wang, and Dinesh C Soares, and Hongping Jin, and Haran Sivakumaran, and Min-Hsuan Lin, and Kirsten Spann, and Catherine M Abbott, and David Harrich
Interaction between Reverse Transcriptase and Integrase Is Required for Reverse Transcription during HIV-1 Replication.
December 2015, Journal of virology,
Dongsheng Li, and Ting Wei, and Daniel J Rawle, and Fangyun Qin, and Rui Wang, and Dinesh C Soares, and Hongping Jin, and Haran Sivakumaran, and Min-Hsuan Lin, and Kirsten Spann, and Catherine M Abbott, and David Harrich
Oxazole-Benzenesulfonamide Derivatives Inhibit HIV-1 Reverse Transcriptase Interaction with Cellular eEF1A and Reduce Viral Replication.
June 2019, Journal of virology,
Dongsheng Li, and Ting Wei, and Daniel J Rawle, and Fangyun Qin, and Rui Wang, and Dinesh C Soares, and Hongping Jin, and Haran Sivakumaran, and Min-Hsuan Lin, and Kirsten Spann, and Catherine M Abbott, and David Harrich
Augmentation of reverse transcription by integrase through an interaction with host factor, SIP1/Gemin2 Is critical for HIV-1 infection.
November 2009, PloS one,
Dongsheng Li, and Ting Wei, and Daniel J Rawle, and Fangyun Qin, and Rui Wang, and Dinesh C Soares, and Hongping Jin, and Haran Sivakumaran, and Min-Hsuan Lin, and Kirsten Spann, and Catherine M Abbott, and David Harrich
HIV-1 Uncoating and Reverse Transcription Require eEF1A Binding to Surface-Exposed Acidic Residues of the Reverse Transcriptase Thumb Domain.
March 2018, mBio,
Dongsheng Li, and Ting Wei, and Daniel J Rawle, and Fangyun Qin, and Rui Wang, and Dinesh C Soares, and Hongping Jin, and Haran Sivakumaran, and Min-Hsuan Lin, and Kirsten Spann, and Catherine M Abbott, and David Harrich
Isolated HIV-1 core is active for reverse transcription.
October 2007, Retrovirology,
Dongsheng Li, and Ting Wei, and Daniel J Rawle, and Fangyun Qin, and Rui Wang, and Dinesh C Soares, and Hongping Jin, and Haran Sivakumaran, and Min-Hsuan Lin, and Kirsten Spann, and Catherine M Abbott, and David Harrich
Tat is required for efficient HIV-1 reverse transcription.
March 1997, The EMBO journal,
Dongsheng Li, and Ting Wei, and Daniel J Rawle, and Fangyun Qin, and Rui Wang, and Dinesh C Soares, and Hongping Jin, and Haran Sivakumaran, and Min-Hsuan Lin, and Kirsten Spann, and Catherine M Abbott, and David Harrich
HIV Tat/P-TEFb Interaction: A Potential Target for Novel Anti-HIV Therapies.
April 2018, Molecules (Basel, Switzerland),
Copied contents to your clipboard!