Biochemical and virological analysis of the 18-residue C-terminal tail of HIV-1 integrase.

Abstract:

BACKGROUND:The 18 residue tail abutting the SH3 fold that comprises the heart of the C-terminal domain is the only part of HIV-1 integrase yet to be visualized by structural biology. To ascertain the role of the tail region in integrase function and HIV-1 replication, a set of deletion mutants that successively lacked three amino acids was constructed and analyzed in a variety of biochemical and virus infection assays. HIV-1/2 chimers, which harbored the analogous 23-mer HIV-2 tail in place of the HIV-1 sequence, were also studied. Because integrase mutations can affect steps in the replication cycle other than integration, defective mutant viruses were tested for integrase protein content and reverse transcription in addition to integration. The F185K core domain mutation, which increases integrase protein solubility, was furthermore analyzed in a subset of mutants. RESULTS:Purified proteins were assessed for in vitro levels of 3' processing and DNA strand transfer activities whereas HIV-1 infectivity was measured using luciferase reporter viruses. Deletions lacking up to 9 amino acids (1-285, 1-282, and 1-279) displayed near wild-type activities in vitro and during infection. Further deletion yielded two viruses, HIV-1(1-276) and HIV-1(1-273), that displayed approximately two and 5-fold infectivity defects, respectively, due to reduced integrase function. Deletion mutant HIV-1(1-270) and the HIV-1/2 chimera were non-infectious and displayed approximately 3 to 4-fold reverse transcription in addition to severe integration defects. Removal of four additional residues, which encompassed the C-terminal beta strand of the SH3 fold, further compromised integrase incorporation into virions and reverse transcription. CONCLUSION:HIV-1(1-270), HIV-1(1-266), and the HIV-1/2 chimera were typed as class II mutant viruses due to their pleiotropic replication defects. We speculate that residues 271-273 might play a role in mediating the known integrase-reverse transcriptase interaction, as their removal unveiled a reverse transcription defect. The F185K mutation reduced the in vitro activities of 1-279 and 1-276 integrases by about 25%. Mutant proteins 1-279/F185K and 1-276/F185K are therefore highlighted as potential structural biology candidates, whereas further deleted tail variants (1-273/F185K or 1-270/F185K) are less desirable due to marginal or undetectable levels of integrase function.

journal_name

Retrovirology

journal_title

Retrovirology

authors

Dar MJ,Monel B,Krishnan L,Shun MC,Di Nunzio F,Helland DE,Engelman A

doi

10.1186/1742-4690-6-94

subject

Has Abstract

pub_date

2009-10-19 00:00:00

pages

94

issn

1742-4690

pii

1742-4690-6-94

journal_volume

6

pub_type

杂志文章
  • The Infectious Diseases BioBank at King's College London: archiving samples from patients infected with HIV to facilitate translational research.

    abstract::The King's College London (KCL) Infectious Diseases BioBank opened in 2007 and collects peripheral venous blood (PVB) from individuals infected with pathogens including human immunodeficiency virus (HIV). PVBs are fractionated into plasmas, lymphocytes and DNA and are then frozen. All donations are from subjects who h...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/1742-4690-6-98

    authors: Williams R,Mant C,Cason J

    更新日期:2009-11-03 00:00:00

  • Avian myeoloblastosis virus (AMV): only one side of the coin.

    abstract::For many years, scientists and suppliers have refered to AMV-RT as the reverse transcriptase produced by the Avian Myelobalstosis Virus. This manuscript briefly reviews the molecular basis for biological dependence of AMV for the envelope and RT proteins that are produced by its natural helper the Myeloblastosis Assoc...

    journal_title:Retrovirology

    pub_type: 杂志文章,评审

    doi:10.1186/1742-4690-5-49

    authors: Perbal B

    更新日期:2008-06-16 00:00:00

  • Higher levels of Zidovudine resistant HIV in the colon compared to blood and other gastrointestinal compartments in HIV infection.

    abstract:BACKGROUND:The gut-associated lymphoid tissue (GALT) is the largest lymphoid organ infected by human immunodeficiency virus type 1 (HIV-1). It serves as a viral reservoir and host-pathogen interface in infection. This study examined whether different parts of the gut and peripheral blood lymphocytes (PBL) contain diffe...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/1742-4690-7-74

    authors: van Marle G,Church DL,Nunweiler KD,Cannon K,Wainberg MA,Gill MJ

    更新日期:2010-09-13 00:00:00

  • microRNAs in viral oncogenesis.

    abstract::MicroRNAs are a recently discovered class of small noncoding functional RNAs. These molecules mediate post-transcriptional regulation of gene expression in a sequence specific manner. MicroRNAs are now known to be key players in a variety of biological processes and have been shown to be deregulated in a number of can...

    journal_title:Retrovirology

    pub_type: 社论,评审

    doi:10.1186/1742-4690-4-82

    authors: Scaria V,Jadhav V

    更新日期:2007-11-24 00:00:00

  • Identification of the feline foamy virus Bet domain essential for APOBEC3 counteraction.

    abstract:BACKGROUND:APOBEC3 (A3) proteins restrict viral replication by cytidine deamination of viral DNA genomes and impairing reverse transcription and integration. To escape this restriction, lentiviruses have evolved the viral infectivity factor (Vif), which binds A3 proteins and targets them for proteolytic degradation. In...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/1742-4690-10-76

    authors: Lukic DS,Hotz-Wagenblatt A,Lei J,Räthe AM,Mühle M,Denner J,Münk C,Löchelt M

    更新日期:2013-07-24 00:00:00

  • Utilization of HIV-1 envelope V3 to identify X4- and R5-specific Tat and LTR sequence signatures.

    abstract:BACKGROUND:HIV-1 entry is a receptor-mediated process directed by the interaction of the viral envelope with the host cell CD4 molecule and one of two co-receptors, CCR5 or CXCR4. The amino acid sequence of the third variable (V3) loop of the HIV-1 envelope is highly predictive of co-receptor utilization preference dur...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/s12977-016-0266-9

    authors: Antell GC,Dampier W,Aiamkitsumrit B,Nonnemacher MR,Jacobson JM,Pirrone V,Zhong W,Kercher K,Passic S,Williams JW,Schwartz G,Hershberg U,Krebs FC,Wigdahl B

    更新日期:2016-05-03 00:00:00

  • OVEX1, a novel chicken endogenous retrovirus with sex-specific and left-right asymmetrical expression in gonads.

    abstract:BACKGROUND:In chickens, as in most birds, female gonad morphogenesis is asymmetrical. Gonads appear first rather similarly, but only the left one undergoes full differentiation and gives rise to a functional ovary. The right gonad, in which the cortex does not develop, remains restricted to the medulla and finally regr...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/1742-4690-6-59

    authors: Carré-Eusèbe D,Coudouel N,Magre S

    更新日期:2009-06-17 00:00:00

  • Blockade of chemokine-induced signalling inhibits CCR5-dependent HIV infection in vitro without blocking gp120/CCR5 interaction.

    abstract:BACKGROUND:Cellular infection with human immunodeficiency virus (HIV) both in vitro and in vivo requires a member of the chemokine receptor family to act as a co-receptor for viral entry. However, it is presently unclear to what extent the interaction of HIV proteins with chemokine receptors generates intracellular sig...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/1742-4690-2-23

    authors: Grainger DJ,Lever AM

    更新日期:2005-04-04 00:00:00

  • Stabilization of the β-hairpin in Mason-Pfizer monkey virus capsid protein- a critical step for infectivity.

    abstract:BACKGROUND:Formation of a mature core is a crucial event for infectivity of retroviruses such as Mason-Pfizer monkey virus (M-PMV). The process is triggered by proteolytic cleavage of the polyprotein precursor Gag, which releases matrix, capsid (CA), and nucleocapsid proteins. Once released, CA assembles to form a matu...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/s12977-014-0094-8

    authors: Obr M,Hadravová R,DoleŽal M,KříŽová I,Papoušková V,Zídek L,Hrabal R,Ruml T,Rumlová M

    更新日期:2014-10-30 00:00:00

  • CD45 immunoaffinity depletion of vesicles from Jurkat T cells demonstrates that exosomes contain CD45: no evidence for a distinct exosome/HIV-1 budding pathway.

    abstract::The presence of relatively high levels of cellular protein contamination in density-purified virion preparations is a confounding factor in biochemical analyses of HIV and SIV produced from hematopoietic cells. A major source of this contamination is from vesicles, either microvesicles or exosomes, that have similar p...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/1742-4690-5-64

    authors: Coren LV,Shatzer T,Ott DE

    更新日期:2008-07-16 00:00:00

  • The co-receptor signaling model of HIV-1 pathogenesis in peripheral CD4 T cells.

    abstract::HIV-mediated CD4 depletion is the hallmark of AIDS and is the most reliable predictor of disease progression. While HIV replication is associated with CD4 depletion in general, plasma viremia by itself predicts the rate of CD4 loss only minimally in untreated patients. To resolve this paradox, I hypothesize the existe...

    journal_title:Retrovirology

    pub_type: 社论

    doi:10.1186/1742-4690-6-41

    authors: Wu Y

    更新日期:2009-05-01 00:00:00

  • TLR8 regulation of LILRA3 in monocytes is abrogated in human immunodeficiency virus infection and correlates to CD4 counts and virus loads.

    abstract:BACKGROUND:LILRA3 is an immunostimulatory molecule which can conditionally induce the proliferation of cytotoxic cells. LILRA3 has a deletion genotype which is associated with multiple immune disorders. In this study, we wanted to analyze the regulation of LILRA3 and its significance in the context of HIV infection. R...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/s12977-016-0248-y

    authors: Low HZ,Ahrenstorf G,Pommerenke C,Habermann N,Schughart K,Ordóñez D,Stripecke R,Wilk E,Witte T

    更新日期:2016-03-12 00:00:00

  • Inhibition of Tat activity by the HEXIM1 protein.

    abstract:BACKGROUND:The positive transcription elongation factor b (P-TEFb) composed by CDK9/CyclinT1 subunits is a dedicated co-factor of HIV transcriptional transactivator Tat protein. Transcription driven by the long terminal repeat (LTR) of HIV involves formation of a quaternary complex between P-TEFb, Tat and the TAR eleme...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/1742-4690-2-42

    authors: Fraldi A,Varrone F,Napolitano G,Michels AA,Majello B,Bensaude O,Lania L

    更新日期:2005-07-02 00:00:00

  • Virus-producing cells determine the host protein profiles of HIV-1 virion cores.

    abstract:BACKGROUND:Upon HIV entry into target cells, viral cores are released and rearranged into reverse transcription complexes (RTCs), which support reverse transcription and also protect and transport viral cDNA to the site of integration. RTCs are composed of viral and cellular proteins that originate from both target and...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/1742-4690-9-65

    authors: Santos S,Obukhov Y,Nekhai S,Bukrinsky M,Iordanskiy S

    更新日期:2012-08-13 00:00:00

  • Digital PCR as a tool to measure HIV persistence.

    abstract::Although antiretroviral therapy is able to suppress HIV replication in infected patients, the virus persists and rebounds when treatment is stopped. In order to find a cure that can eradicate the latent reservoir, one must be able to quantify the persisting virus. Traditionally, HIV persistence studies have used real-...

    journal_title:Retrovirology

    pub_type: 杂志文章,评审

    doi:10.1186/s12977-018-0399-0

    authors: Rutsaert S,Bosman K,Trypsteen W,Nijhuis M,Vandekerckhove L

    更新日期:2018-01-30 00:00:00

  • HLA-B*35 as a new marker for susceptibility to human T-cell lymphotropic virus type 1 (HTLV-1) Associated Myelopathy/Tropical Spastic Paraparesis (HAM/TSP) in patients living in Argentina.

    abstract:BACKGROUND:Human T-cell lymphotropic virus type 1 (HTLV-1) is the etiological agent of HTLV associated myelopathy/ Tropical Spastic Paraparesis (HAM/TSP) and Adult T cell leukemia/lymphoma (ATLL), in around 2-5% of the infected individuals. Host genetic background might play a role in disease progression. Several previ...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/s12977-020-00536-y

    authors: Benencio P,Fraile Gonzalez SA,Ducasa N,Page K,Berini CA,Biglione MM

    更新日期:2020-09-03 00:00:00

  • TNPO3 protects HIV-1 replication from CPSF6-mediated capsid stabilization in the host cell cytoplasm.

    abstract:BACKGROUND:Despite intensive investigation the mechanism by which HIV-1 reaches the host cell nucleus is unknown. TNPO3, a karyopherin mediating nuclear entry of SR-proteins, was shown to be required for HIV-1 infectivity. Some investigators have reported that TNPO3 promotes HIV-1 nuclear import, as would be expected f...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/1742-4690-10-20

    authors: De Iaco A,Santoni F,Vannier A,Guipponi M,Antonarakis S,Luban J

    更新日期:2013-02-15 00:00:00

  • The inhibition of microRNAs by HIV-1 Tat suppresses beta catenin activity in astrocytes.

    abstract:BACKGROUND:Long term infection with HIV-1, even in the context of therapy, leads to chronic health problems including an array of neurocognitive dysfunctions. The viral Tat protein has previously been implicated in neuropathogenesis through its effect on astrocytes. Tat has also been shown to inhibit the biogenesis of ...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/s12977-016-0256-y

    authors: Sardo L,Vakil PR,Elbezanti W,El-Sayed A,Klase Z

    更新日期:2016-04-08 00:00:00

  • HIV-1 envelope replication and α4β7 utilization among newly infected subjects and their corresponding heterosexual partners.

    abstract:BACKGROUND:Previous studies suggest that active selection limits the number of HIV-1 variants acquired by a newly infected individual from the diverse variants circulating in the transmitting partner. We compared HIV-1 envelopes from 9 newly infected subjects and their linked transmitting partner to explore potential m...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/1742-4690-10-162

    authors: Pena-Cruz V,Etemad B,Chatziandreou N,Nyein PH,Stock S,Reynolds SJ,Laeyendecker O,Gray RH,Serwadda D,Lee SJ,Quinn TC,Sagar M

    更新日期:2013-12-26 00:00:00

  • The distribution of insertionally polymorphic endogenous retroviruses in breast cancer patients and cancer-free controls.

    abstract:BACKGROUND:Integration of retroviral DNA into a germ cell can result in a provirus that is transmitted vertically to the host's offspring. In humans, such endogenous retroviruses (HERVs) comprise >8% of the genome. The HERV-K(HML-2) proviruses consist of ~90 elements related to mouse mammary tumor virus, which causes b...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/s12977-014-0062-3

    authors: Wildschutte JH,Ram D,Subramanian R,Stevens VL,Coffin JM

    更新日期:2014-08-12 00:00:00

  • How to engage Cofilin.

    abstract::In HIV-infected people, resting CD4+ T cells are the main reservoir of latent virus and the reason for the failure of drug therapy to cure HIV infection. Still, we do not have a complete understanding of the factors regulating HIV replication in these cells. A recent paper in Cell describes a new trick that the virus ...

    journal_title:Retrovirology

    pub_type: 杂志文章,评审

    doi:10.1186/1742-4690-5-85

    authors: Bukrinsky M

    更新日期:2008-09-22 00:00:00

  • Contamination of clinical specimens with MLV-encoding nucleic acids: implications for XMRV and other candidate human retroviruses.

    abstract::Efforts to assess the prevalence of xenotropic murine leukemia virus-related virus (XMRV) in patients with prostate cancer and chronic fatigue syndrome have relied heavily on PCR-based testing of clinical samples and have yielded widely divergent findings. This week in Retrovirology, reports from four independent rese...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/1742-4690-7-112

    authors: Smith RA

    更新日期:2010-12-20 00:00:00

  • World AIDS Day 2007: AIDS at 26, are we there yet?

    abstract::This editorial comments on selected progress made in combating the acquired immune deficiency syndrome (AIDS) after 26 years and some of the remaining challenges. ...

    journal_title:Retrovirology

    pub_type: 社论

    doi:10.1186/1742-4690-4-86

    authors: Jeang KT

    更新日期:2007-12-01 00:00:00

  • DBR1 siRNA inhibition of HIV-1 replication.

    abstract:BACKGROUND:HIV-1 and all retroviruses are related to retroelements of simpler organisms such as the yeast Ty elements. Recent work has suggested that the yeast retroelement Ty1 replicates via an unexpected RNA lariat intermediate in cDNA synthesis. The putative genomic RNA lariat intermediate is formed by a 2'-5' phosp...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/1742-4690-2-63

    authors: Ye Y,De Leon J,Yokoyama N,Naidu Y,Camerini D

    更新日期:2005-10-18 00:00:00

  • MDM2 is a novel E3 ligase for HIV-1 Vif.

    abstract::The human immunodeficiency virus type 1 (HIV-1) Vif plays a crucial role in the viral life cycle by antagonizing a host restriction factor APOBEC3G (A3G). Vif interacts with A3G and induces its polyubiquitination and subsequent degradation via the formation of active ubiquitin ligase (E3) complex with Cullin5-ElonginB...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/1742-4690-6-1

    authors: Izumi T,Takaori-Kondo A,Shirakawa K,Higashitsuji H,Itoh K,Io K,Matsui M,Iwai K,Kondoh H,Sato T,Tomonaga M,Ikeda S,Akari H,Koyanagi Y,Fujita J,Uchiyama T

    更新日期:2009-01-07 00:00:00

  • Generation and validation of a highly sensitive bioluminescent HIV-1 reporter vector that simplifies measurement of virus release.

    abstract:BACKGROUND:The continued persistence of HIV-1 as a public health concern due to the lack of a cure calls for the development of new tools for studying replication of the virus. Here, we used NanoLuc, a small and extremely bright luciferase protein, to develop an HIV-1 bioluminescent reporter virus that simplifies funct...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/s12977-020-00521-5

    authors: Kirui J,Freed EO

    更新日期:2020-05-19 00:00:00

  • Preferences for the selection of unique tRNA primers revealed from analysis of HIV-1 replication in peripheral blood mononuclear cells.

    abstract:BACKGROUND:All human immunodeficiency virus (HIV-1) uses a host tRNALys,3 as the primer for reverse transcription. The tRNALys,3 is bound to a region on the HIV-1 genome, the primer-binding site (PBS), that is complementary to the 18 terminal nucleotides of tRNALys,3. How HIV-1 selects the tRNA from the intracellular m...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/1742-4690-2-21

    authors: Moore-Rigdon KL,Kosloff BR,Kirkman RL,Morrow CD

    更新日期:2005-03-24 00:00:00

  • The role of cyclin D2 and p21/waf1 in human T-cell leukemia virus type 1 infected cells.

    abstract:BACKGROUND:The human T-cell leukemia virus type 1 (HTLV-1) Tax protein indirectly influences transcriptional activation, signal transduction, cell cycle control, and apoptosis. The function of Tax primarily relies on protein-protein interactions. We have previously shown that Tax upregulates the cell cycle checkpoint p...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/1742-4690-1-6

    authors: Kehn K,Deng L,de la Fuente C,Strouss K,Wu K,Maddukuri A,Baylor S,Rufner R,Pumfery A,Bottazzi ME,Kashanchi F

    更新日期:2004-04-13 00:00:00

  • Modulation of HIV-1 Gag NC/p1 cleavage efficiency affects protease inhibitor resistance and viral replicative capacity.

    abstract:BACKGROUND:Mutations in the substrate of HIV-1 protease, especially changes in the NC/p1 cleavage site, can directly contribute to protease inhibitor (PI) resistance and also compensate for defects in viral replicative capacity (RC) due to a drug resistant protease. These NC/p1 changes are known to enhance processing o...

    journal_title:Retrovirology

    pub_type: 杂志文章

    doi:10.1186/1742-4690-9-29

    authors: van Maarseveen NM,Andersson D,Lepšík M,Fun A,Schipper PJ,de Jong D,Boucher CA,Nijhuis M

    更新日期:2012-04-01 00:00:00

  • STLV-1 as a model for studying HTLV-1 infection.

    abstract::Few years after HTLV-1 identification and isolation in humans, STLV-1, its simian counterpart, was discovered. It then became clear that STLV-1 is present almost in all simian species. Subsequent molecular epidemiology studies demonstrated that, apart from HTLV-1 subtype A, all human subtypes have a simian homolog. As...

    journal_title:Retrovirology

    pub_type: 杂志文章,评审

    doi:10.1186/s12977-019-0503-0

    authors: Jégado B,Kashanchi F,Dutartre H,Mahieux R

    更新日期:2019-12-16 00:00:00