Method: Briefly, CA structures were self-assembled using a mixture of CA A204C/A92E (for cross-linking and increased solubility, respectively) and AF647-labelled CA K158C and then captured on the surface of a coverslip.
Figure: (b) Dissociation constants (KD's) of Atto488-ATP binding to self-assembled CA A204C/A92E cones determined by TIRF microscopy.
Characterization of HIV-1 uncoating in human microglial cell lines.
Result: Finally, A92E mutant virus had an uncoating half-life of 62 min that was not statistically different from the 52 min for wildtype.
Result: Some alterations in completion of reverse transcription were observed, with A92E seeming to reverse transcribe at the greatest rate and E45A at the slowest rate.
Result: We previously examined the uncoating of a panel of CA mutant viruses in OMK cells and found that the N74D, A92E, and E45A mutations altered the rate of uncoating compared to wildtype.
Figure: d The A92E mutation did not significantly alter the rate of uncoating among seven independent experiments.
Discussion: A92E
Cyclophilin A Prevents HIV-1 Restriction in Lymphocytes by Blocking Human TRIM5alpha Binding to the Viral Core.
Abstract: Disruption of CypA-capsid interactions fail to affect HIV-1-A92E/G94D infection, correlating with the loss of TRIM5alphahu binding to HIV-1-A92E/G94D cores.
Introduction: In agreement with our hypothesis that CypA protects the core from TRIM5alphahu, infection of PBMCs and CD4+ T cells by HIV-1-A92E viruses was insensitive to the disruption of CypA-capsid interactions.
Introduction: Infection of HIV-1-A92E/G94D viruses was not affected by disruption of CypA-capsid interactions, which correlates with the loss of TRIM5alphahu binding to HIV-1 cores bearing the A92E/
MxB sensitivity of HIV-1 is determined by a highly variable and dynamic capsid surface.
Result: Importantly, the A92E CA mutation also rescued MxB sensitivity in the absence of CypA protein, after shRNA-mediated CypA depletion, and after CypA inhibition with CsA (Figure 3H and I).
Result: In fact, adding A92E to HIV-1 CA P90A rescued sensitivity to MxB.
Result: The A92E mutation was identified by selection of HIV-1 replicating mutants during CypA inhibition with CsA.
Result: These data demonstrate that, for this particular HIV-1 isolate (HIV-1 R9), it is not CypA recruitment per se that is required for MxB sensitivity, but rather the conformational effect of CypA recruitment, likely the CypA mediated stabilisation of the CypA binding loop, which is recapitulated by the A92E and PMID: 30947724
2019
Retrovirology
Abstract: In normal HeLa-P4 cells, infection with HIV-1 bearing the A92E substitution in CA is inhibited by endogenous CypA and was inhibited to the same extent by expression of CypA mutants in CypA-null HeLa-P4 cells.
Result: In the following sections we present results from infection assays in (1) HeLa-P4 cells (using HIV-1 with CA A92E mutation) and (2) Jurkat cells.
Result: The HIV-1 capsid escape mutant A92E arises during passage of HIV-1 in HeLa-P4 cells during CypA inhibition and is poorly infective in cells expressing high CypA le
Figure: a Analysis of the extent of HIV-1 infection of the indicated cell lines by the A92E CA mutant in the presence and absence of CsA.
Multiple Pathways To Avoid Beta Interferon Sensitivity of HIV-1 by Mutations in Capsid.
Result: 10H and I), suggesting that the G116R mutation augmented the infectivity of both the RGDA/Q112D+A92E and the RGDA/Q112D+G94D viruses.
Result: Consistent with previous studies, infections of the NL4-3 G94D and A92E viruses were enhanced by CypA knockout or CsA treatment in HeLa cells.
Result: The RGDA/Q112D+A92E/G116R virus exhibited infectivity significantly higher than that of the RGDA/Q112D+Result: We examined whether the G116R mutation augments the infectivity of the RGDA/Q112D+A92E virus.
Quenching protein dynamics interferes with HIV capsid maturation.
Method: A helical tube of mature-like CA(A92E)-SP1 NL4-3 was constructed by docking the MDFF-derived hexameric structure (PDB: 3J34), using Chimera, into the density of hexamer of tubes with (-8, 14) symmetry.
Method: Construction of an immature-like lattice of CA(A92E)-SP1 was performed by docking monomeric HIV-1 CTD (PDB: 2KOD) into the cryo-EM density derived from authentic virions (EMDB: 2706).
Method: Full-length CA(A92E)-SP1 NL4-3 proteins (3.95 mg/ml) were diluted to 2.2 mg/ml in high salt buffer (1 M NaCl, 50 mM Tris pH 8.0) and incubated at 37 C for 1 h for tubular assembly.
Identification of capsid mutations that alter the rate of HIV-1 uncoating in infected cells.
Abstract: We found that p24(CA) mutations can significantly increase (A92E), delay (E45A and N74D), or have no effect (G94D) on the rate of uncoating and that these alterations are not due to changes in reverse transcription.
Abstract: Remarkably, the CypA loop dynamics of wild-type CA HXB2 assembly is significantly attenuated upon CypA binding, and the dynamics profiles of the A92E and G94D CypA escape mutants closely resemble that of wild-type CA assembly in complex with CypA.
Abstract: Through the analysis of backbone (1)H-(15)N and (1)H-(13)C dipolar tensors and peak intensities from 3D MAS NMR spectra of wild-type and the A92E and G94D CypA escape mutants, we demonstrate that assembled CA is dynamic, particularly in loop regions.
Cyclophilin A promotes HIV-1 reverse transcription but its effect on transduction correlates best with its effect on nuclear entry of viral cDNA.
Abstract: Transduction of certain cell types increases significantly when CypA binding to particular HIV-1 CA mutants, i.e., A92E, is prevented.
Introduction: HIV-1 viruses bearing either the A92E or G94D CA mutation are defective for replication in HeLa or H9 T cells, but disruption of the CypA-CA interaction rescued infectivity of these mutants in these cells.
Introduction: How CypA promotes HIV-1 infection or contributes to the block to A92E or G94D mutant virus replication in certain cell lines is not well understood.
Introduction: Interestingly, CA mutations N74D and A105T
HIV mutation literature information.
PMID: 
2021
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