Molecular mimicry of human tRNALys anti-codon domain by HIV-1 RNA genome facilitates tRNA primer annealing

The HIV-1 integrase enzyme (IN) plays a critical role in the viral life cycle by integrating the reverse-transcribed viral DNA into the host chromosome. This function of IN has been well studied, and the knowledge gained has informed the design of small molecule inhibitors that now form key components of antiretroviral therapy regimens. Recent discoveries unveiled that IN has an under-studied yet equally vital second function in human immunodeficiency virus type 1 (HIV-1) replication. This involves IN binding to the viral RNA genome in virions, which is necessary for proper virion maturation and morphogenesis. Inhibition of IN binding to the viral RNA genome results in mislocalization of the viral genome inside the virus particle, and its premature exposure and degradation in target cells. The roles of IN in integration and virion morphogenesis share a number of common elements, including interaction with viral nucleic acids and assembly of higher-order IN multimers. Herein we describe these two functions of IN within the context of the HIV-1 life cycle, how IN binding to the viral genome is coordinated by the major structural protein, Gag, and discuss the value of targeting the second role of IN in virion morphogenesis.

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Selection of Retroviral Reverse Transcription Primer Is Coordinated with tRNA Biogenesis

Journal of Virology ◽

10.1128/jvi.77.16.8695-8701.2003 ◽

2003 ◽

Vol 77 (16) ◽

pp. 8695-8701 ◽

Cited By ~ 21

Author(s):

Nathan J. Kelly ◽

Matthew T. Palmer ◽

Casey D. Morrow

Keyword(s):

Reverse Transcription ◽

Nuclear Export ◽

Mammalian Cells ◽

Leukemia Virus ◽

Wild Type ◽

Primer Selection ◽

Yeast Trna ◽

Trna Primer ◽

Hiv 1 ◽

Selection Of

ABSTRACT Initiation of retrovirus reverse transcription requires the selection of a tRNA primer from the intracellular milieu. To investigate the features of primer selection, a human immunodeficiency virus type 1 (HIV-1) and a murine leukemia virus (MuLV) were created that require yeast tRNAPhe to be supplied in trans for infectivity. Wild-type yeast tRNAPhe expressed in mammalian cells was transported to the cytoplasm and aminoacylated. In contrast, tRNAPhe without the D loop (tRNAPheD−) was retained within the nucleus and did not complement infectivity of either HIV-1 or MuLV; however, infectivity was restored when tRNAPheD− was directly transfected into the cytoplasm of cells. A tRNAPhe mutant (tRNAPheUUA) that did not have the capacity to be aminoacylated was transported to the cytoplasm and did complement infectivity of both HIV-1 and MuLV, albeit at a level less than that with wild-type tRNAPhe. Collectively, our results demonstrate that the tRNA primer captured by HIV-1 and MuLV occurs after nuclear export of tRNA and supports a model in which primer selection for retroviruses is coordinated with tRNA biogenesis at the intracellular site of protein synthesis.

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How Polypurine Tract Changes in the HIV-1 RNA Genome Can Cause Resistance against the Integrase Inhibitor Dolutegravir

mBio ◽

10.1128/mbio.00006-18 ◽

2018 ◽

Vol 9 (2) ◽

Cited By ~ 6

Author(s):

Atze T. Das ◽

Ben Berkhout

Keyword(s):

Integrase Inhibitor ◽

Polypurine Tract ◽

Rna Genome ◽

Hiv 1

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RNA and Nucleocapsid Are Dispensable for Mature HIV-1 Capsid Assembly

Journal of Virology ◽

10.1128/jvi.00750-15 ◽

2015 ◽

Vol 89 (19) ◽

pp. 9739-9747 ◽

Cited By ~ 8

Author(s):

Simone Mattei ◽

Annica Flemming ◽

Maria Anders-Össwein ◽

Hans-Georg Kräusslich ◽

John A. G. Briggs ◽

...

Keyword(s):

Nucleocapsid Protein ◽

Rna Binding ◽

Electron Tomography ◽

Significant Proportion ◽

Dimensional Structure ◽

Capsid Assembly ◽

Interaction Domain ◽

Cryo Electron Tomography ◽

Rna Genome ◽

Hiv 1

ABSTRACTHuman immunodeficiency virus type 1 (HIV-1) is released from infected cells in an immature, noninfectious form in which the structural polyprotein Gag is arranged in a hexameric lattice, forming an incomplete spherical shell. Maturation to the infectious form is mediated by the viral protease, which cleaves Gag at five sites, releasing the CA (capsid) protein, which forms a conical capsid encasing the condensed RNA genome. The pathway of this structural rearrangement is currently not understood, and it is unclear how cone assembly is initiated. RNA represents an integral structural component of retroviruses, and the viral nucleoprotein core has previously been proposed to nucleate mature capsid assembly. We addressed this hypothesis by replacing the RNA-binding NC (nucleocapsid) domain of HIV-1 Gag and the adjacent spacer peptide 2 (SP2) by a leucine zipper (LZ) protein-protein interaction domain [Gag(LZ)] in the viral context. We found that Gag(LZ)-carrying virus [HIV(LZ)] was efficiently released and viral polyproteins were proteolytically processed, though with reduced efficiency. Cryo-electron tomography revealed that the particles lacked a condensed nucleoprotein and contained an increased proportion of aberrant core morphologies caused either by the absence of RNA or by altered Gag processing. Nevertheless, a significant proportion of HIV(LZ) particles contained mature capsids with the wild-type morphology. These results clearly demonstrate that the nucleoprotein complex is dispensable as a nucleator for mature HIV-1 capsid assembly in the viral context.IMPORTANCEFormation of a closed conical capsid encasing the viral RNA genome is essential for HIV-1 infectivity. It is currently unclear what viral components initiate and regulate the formation of the capsid during virus morphogenesis, but it has been proposed that the ribonucleoprotein complex plays a role. To test this, we prepared virus-like particles lacking the viral nucleocapsid protein and RNA and analyzed their three-dimensional structure by cryo-electron tomography. While most virions displayed an abnormal morphology under these conditions, some particles showed a normal mature morphology with closed conical capsids. These data demonstrate that the presence of RNA and the nucleocapsid protein is not required for the formation of a mature, cone-shaped HIV-1 capsid.

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Unexpected Molecular Mimicry among Peptides MHC Class II, Blood-Clotting Factor X, and HIV-1 Envelope Glycoprotein GP120

Thrombosis Research ◽

10.1016/s0049-3848(99)00245-5 ◽

2000 ◽

Vol 98 (4) ◽

pp. 343-346 ◽

Cited By ~ 1

Author(s):

Garson H Tishkoff ◽

Lois T Hunt

Keyword(s):

Mhc Class Ii ◽

Envelope Glycoprotein ◽

Blood Clotting ◽

Molecular Mimicry ◽

Class Ii ◽

Clotting Factor ◽

Factor X ◽

Glycoprotein Gp120 ◽

Hiv 1 ◽

Blood Clotting Factor

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Mutations in the U5 Region Adjacent to the Primer Binding Site Affect tRNA Cleavage by Human Immunodeficiency Virus Type 1 Reverse Transcriptase In Vivo

Journal of Virology ◽

10.1128/jvi.02611-06 ◽

2007 ◽

Vol 82 (2) ◽

pp. 719-727 ◽

Cited By ~ 11

Author(s):

Jangsuk Oh ◽

Mary Jane McWilliams ◽

John G. Julias ◽

Stephen H. Hughes

Keyword(s):

Human Immunodeficiency Virus ◽

Reverse Transcriptase ◽

Binding Site ◽

Rnase H ◽

Primer Binding Site ◽

Cleavage Specificity ◽

Immunodeficiency Virus ◽

Trna Primer ◽

Hiv 1

ABSTRACT In retroviruses, the first nucleotide added to the tRNA primer defines the end of the U5 region in the right long terminal repeat, and the subsequent removal of this tRNA primer by RNase H exactly defines the U5 end of the linear double-stranded DNA. In most retroviruses, the entire tRNA is removed by RNase H cleavage at the RNA/DNA junction. However, the RNase H domain of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase cleaves the tRNA 1 nucleotide from the RNA/DNA junction at the U5/primer binding site (PBS) junction, which leaves an rA residue at the U5 terminus. We made sequence changes at the end of the U5 region adjacent to the PBS in HIV-1 to determine whether such changes affect the specificity of tRNA primer cleavage by RNase H. In some of the mutants, RNase H usually removed the entire tRNA, showing that the cleavage specificity was shifted by 1 nucleotide. This result suggests that the tRNA cleavage specificity of the HIV-1 RNase domain H depends on sequences in U5.

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