HIV-1 Tat Length: Comparative and Functional Considerations

Mele, Anthony R.; Marino, Jamie; Dampier, Will; Wigdahl, Brian; Nonnemacher, Michael R.

doi:10.3389/fmicb.2020.00444

Cited by 11 publications

(7 citation statements)

References 51 publications

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“…Previously, we confirmed that ultrapure Tat protein or short PTD-Tat (47-57 amino acids) was entering into neuronal cells or nucleus as early as after 40 minutes post-treatment [37,38]. Our and other research groups investigated features and characteristics of Tat protein to achieve an efficient transduction and to avoid degradation or cleavage prior its transportation into nucleus [39][40][41][42][43].…”

Section: Mitochondrial Dna Damage In Human Primary Neuronal Cells Induced By Recombinant Hiv-1 Tat Peptide or Adenoviral Vprsupporting

confidence: 58%

HIV-1 and HIV-1-Tat Induce Mitochondrial DNA Damage in Human Neurons

et al. 2020

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Introduction: Mitochondrial dysregulation is a key event in HIV-1 infection. Recent studies have suggested that age-related neurodegenerative disorders are associated with increased mitochondrial DNA (mtDNA) damage. As accelerated ageing was found in HIV-1 patients, we hypothesized that HIV-1 infection or HIV-1 proteins can lead to mtDNA damage. Unrepaired mtDNA impairs mitochondrial function, which can lead to oxidative stress and cell death. Investigations of mechanisms of mtDNA damage are limited by the lack of available human models.

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Section: Mitochondrial Dna Damage In Human Primary Neuronal Cells Induced By Recombinant Hiv-1 Tat Peptide or Adenoviral Vprsupporting

confidence: 58%

HIV-1 and HIV-1-Tat Induce Mitochondrial DNA Damage in Human Neurons

et al. 2020

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“…There are multiple splice variants of tat, and the most studied version is a truncated tat 1–86. A recent meta-analysis uncovered that 40% of the 973 studies examining Tat used the 1–86 variant, while only 15.5% used Tat 1–101, which is the most common splice variant observed in people infected with HIV-1 subtype B [ 97 ]. This suggests that one should exercise some caution when considering studies using tat 1–86 or other truncated forms of tat.…”

Section: Synapodendritic Damage and Neuronal Activity Changes In Hand Experimental Modelsmentioning

confidence: 99%

“…This suggests that one should exercise some caution when considering studies using tat 1–86 or other truncated forms of tat. Indeed, there are functional differences reported between tat splice variants that may contribute to synaptodendritic injury in HAND, and common tat-transgenic animal models typically do not express the full-length tat 1–101 [ 97 ]. That said, tat 1–86 exposure in animal models does affect dendritic spines and neuronal function in brain regions including the hippocampus, cortex, and striatum.…”

Section: Synapodendritic Damage and Neuronal Activity Changes In Hand Experimental Modelsmentioning

confidence: 99%

Mechanisms of neuronal dysfunction in HIV-associated neurocognitive disorders

Irollo

Luchetta

et al. 2021

Cell. Mol. Life Sci.

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HIV-associated neurocognitive disorder (HAND) is characterized by cognitive and behavioral deficits in people living with HIV. HAND is still common in patients that take antiretroviral therapies, although they tend to present with less severe symptoms. The continued prevalence of HAND in treated patients is a major therapeutic challenge, as even minor cognitive impairment decreases patient’s quality of life. Therefore, modern HAND research aims to broaden our understanding of the mechanisms that drive cognitive impairment in people with HIV and identify promising molecular pathways and targets that could be exploited therapeutically. Recent studies suggest that HAND in treated patients is at least partially induced by subtle synaptodendritic damage and disruption of neuronal networks in brain areas that mediate learning, memory, and executive functions. Although the causes of subtle neuronal dysfunction are varied, reversing synaptodendritic damage in animal models restores cognitive function and thus highlights a promising therapeutic approach. In this review, we examine evidence of synaptodendritic damage and disrupted neuronal connectivity in HAND from clinical neuroimaging and neuropathology studies and discuss studies in HAND models that define structural and functional impairment of neurotransmission. Then, we report molecular pathways, mechanisms, and comorbidities involved in this neuronal dysfunction, discuss new approaches to reverse neuronal damage, and highlight current gaps in knowledge. Continued research on the manifestation and mechanisms of synaptic injury and network dysfunction in HAND patients and experimental models will be critical if we are to develop safe and effective therapies that reverse subtle neuropathology and cognitive impairment.

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“…The near-perfect accuracy, PPV and NPV for this analysis is consistent with a protease cleavage site between residues 57-58, which is also an exon boundary, and previous observations support similar truncation tolerance for longer trucations. [16][17][18]34,35 However, almost any missense mutation from S46-E86 was tolerated, which is not consistent with the truncation mutants tolerated up to R57. The region of difference (S46-R57) contains a nuclear localization motif, and binding sites for Importinb, P53, and EGR.…”

Section: Intragenic Epistasis Of Tat Double Mutantsmentioning

confidence: 81%

“…[16][17][18] Truncations mutants of less than 58 amino acids from the start Met were considered true negatives, while those 58 amino acids or longer were considered true positives. [16][17][18] Although not designed, due to errors in synthetic oligonucleotide synthesis we observed barcodes for Tat truncation mutants less than 58 amino acids long (n = 70), which were expected to be negatives, and truncation mutants that were longer than 58 amino acids (n = 8) which were expected to be positives with wild type activity (Supplementary File S3). Comparing the Tat mutant activities in LentiX293T to the true controls produced perfect GigaAssay performance statistics for: accuracy = 1.0; sensitivity = 1.0, specificity = 1.0; PPV = 1.0; and NPV = 1.0 (Fig.…”

Section: Gigaassay Performance Verificationmentioning

confidence: 99%

GigaAssay – a high-throughput assay system for molecular functions and cell processes

Schiller

Benjamin

Giacoletto

et al. 2021

Preprint

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High-throughput assay systems have had a disproportionally large impact on uncovering how cells function, as well as how misregulation can lead to disease. However, no high-throughput assay systems have been developed to systematically address how mutations impact molecular functions or cell processes in human cells. This is arguably one of the most critical assays because human pathology and treatment are largely based upon molecular functions. To address this challenge, herein we engineered, developed, and tested the first modular high-throughput molecular function assay system. Note that this is not a selection lethality screen! This “GigaAssay” single cell / one-pot assay system was adapted to study how variants impact HIV Tat-driven transactivation of a green fluorescent protein (GFP) reporter. We assayed all 1,615 Tat single and 3,429 double amino acid substitutions with no single mutant dropout. Each mutant was assayed with replicate observations in LentiX293T and Jurkat cells with an average of 100s of separately barcoded cDNA molecules and cell groups for each mutant. Each mutant had ~2,000X-90,000X sequencing coverage to measure its transcriptional activity and had p value ranging as low as 10-271. Five independent assay performance assessments with benchmark data, individually tested clones, and replicate comparisons all indicate exceptional reproducibility, accuracy, and robustness. The shortcomings of alanine scanning mutagenesis and protein truncation studies are revealed by including exhaustive substitution tolerance and intragenic epistasis in the typical structure/function analysis(structure/function/tolerance/epistasis). This flexible and extensible technology enables a far more comprehensive holistic view of protein molecular function and yet with a highly simplified single-pot assay.

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HIV-1 Tat Length: Comparative and Functional Considerations

Cited by 11 publications

References 51 publications

HIV-1 and HIV-1-Tat Induce Mitochondrial DNA Damage in Human Neurons

HIV-1 and HIV-1-Tat Induce Mitochondrial DNA Damage in Human Neurons

Mechanisms of neuronal dysfunction in HIV-associated neurocognitive disorders

GigaAssay – a high-throughput assay system for molecular functions and cell processes

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