Structural and functional insights into IκB-α/HIV-1 Tat interaction

Vitagliano, Luigi; Fiume, Giuseppe; Scognamiglio, Pasqualina Liana; Doti, Nunzianna; Cannavò, Rosamaria; Puca, Antimina; Pedone, Carlo; Scala, Giuseppe; Quinto, Ileana; Marasco, Daniela

doi:10.1016/j.biochi.2011.05.025

Cited by 20 publications

(22 citation statements)

References 53 publications

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“…It was shown that HIV-1 Tat, a similarly short viral basic protein, is able to bind to several regulator proteins and its binding to the cytosolic IkB-a depends on the arginine-rich region of the protein (residues 48-60) (Vitagliano et al, 2011).…”

Section: Discussionmentioning

confidence: 99%

Immunological and biochemical characterisation of 7ap, a short protein translated from an alternative frame of ORF7 of PRRSV

Olasz

Jankovics

Bálint

et al. 2016

Acta Veterinaria Hungarica

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Sequence analysis revealed a short alternative open reading frame (ORF) named ORF7a within the nucleocapsid gene of genetically divergent porcine reproductive and respiratory syndrome virus (PRRSV) genomes. Alignment of the corresponding protein sequences (named 7ap) revealed substantial heterogeneity among 7aps of different genotypes, though all of them are predicted to be positively charged. Green fluorescent protein and FLAG fusion constructs of ORF7a of the HU-14432/2011 PRRSV demonstrated that 7ap is expressed. 7ap of HU-14432/2011 (Hu7ap) was synthesised chemically, and ELISA experiments revealed that Hu7ap binds strongly to mammalian IgGs. Protein-protein gel retardation assays and complement fixation inhibition suggest that 7aps bind to the CH2 domain of the IgG(Fc) fragment. Cellular localisation and immunological characteristics of PRRSV 7ap may indicate multiple functions including nuclear and cytoplasmic over-tuning of normal cellular processes and immunosuppression.Key words: PRRSV, IgG binding, positive charged short peptide, overlapping ORF, complement fixation inhibition Porcine reproductive and respiratory syndrome virus (PRRSV) is a member of the Arteriviridae family of the Nidovirales order. Since its appearance in the late 1980s PRRSV has remained one of the most costly diseases of the swine industry. PRRSV has a high mutation rate, which led to the evolution of two major genotypes with several subtypes . The nucleotide identity between the two serotypes is only 55-70%. Type

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Section: Discussionmentioning

confidence: 99%

Immunological and biochemical characterisation of 7ap, a short protein translated from an alternative frame of ORF7 of PRRSV

Olasz

Jankovics

Bálint

et al. 2016

Acta Veterinaria Hungarica

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“…The IBTK -shRNA targets the nucleotide sequence from +1534 to +1552 nucleotide of the IBTK α transcript (Ensemble Reference Sequence: ENST00000306270). Lentiviral stocks were produced by transfection of HEK293Tcells, as previously described [28]. Briefly, HEK293Tcells (1 × 10 6 ) were transfected with pCMV-dR8.91 (5 µg) and pCMV-VSVG (5 µg) together with IBTK -shRNA (10 µg) or control-shRNA (10 µg); 48-h post-transfection, cell supernatant was collected.…”

Section: Experimental Sectionsmentioning

confidence: 99%

IBTK Differently Modulates Gene Expression and RNA Splicing in HeLa and K562 Cells

Fiume

Scialdone

Rizzo

et al. 2016

IJMS

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The IBTK gene encodes the major protein isoform IBTKα that was recently characterized as substrate receptor of Cul3-dependent E3 ligase, regulating ubiquitination coupled to proteasomal degradation of Pdcd4, an inhibitor of translation. Due to the presence of Ankyrin-BTB-RCC1 domains that mediate several protein-protein interactions, IBTKα could exert expanded regulatory roles, including interaction with transcription regulators. To verify the effects of IBTKα on gene expression, we analyzed HeLa and K562 cell transcriptomes by RNA-Sequencing before and after IBTK knock-down by shRNA transduction. In HeLa cells, 1285 (2.03%) of 63,128 mapped transcripts were differentially expressed in IBTK-shRNA-transduced cells, as compared to cells treated with control-shRNA, with 587 upregulated (45.7%) and 698 downregulated (54.3%) RNAs. In K562 cells, 1959 (3.1%) of 63128 mapped RNAs were differentially expressed in IBTK-shRNA-transduced cells, including 1053 upregulated (53.7%) and 906 downregulated (46.3%). Only 137 transcripts (0.22%) were commonly deregulated by IBTK silencing in both HeLa and K562 cells, indicating that most IBTKα effects on gene expression are cell type-specific. Based on gene ontology classification, the genes responsive to IBTK are involved in different biological processes, including in particular chromatin and nucleosomal organization, gene expression regulation, and cellular traffic and migration. In addition, IBTK RNA interference affected RNA maturation in both cell lines, as shown by the evidence of alternative 3′- and 5′-splicing, mutually exclusive exons, retained introns, and skipped exons. Altogether, these results indicate that IBTK differently modulates gene expression and RNA splicing in HeLa and K562 cells, demonstrating a novel biological role of this protein.

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“…Cell transfection with the pNF-jB-Luc (pjBLuc) reporter plasmid was carried out as previously described (Fiume et al, 2012;Vitagliano et al, 2011). U251 cells were cultured in a DMEM/Ham's F12 (1:1 ratio) supplemented with 10% heat-inactivated fetal calf serum, 2 mM L-glutamine, 1 mM sodium pyruvate and 1 mM non-essential amino acids, and were transfected with DNA by using FuGENE HD (Roche Diagnostic GmbH, Mannheim, Germany), according to the manufacturer's protocol.…”

Section: Cells Transfection Treatments and Luciferase Assaymentioning

confidence: 99%

Early molecular and behavioral response to lipopolysaccharide in the WAG/Rij rat model of absence epilepsy and depressive-like behavior, involves interplay between AMPK, AKT/mTOR pathways and neuroinflammatory cytokine release

Russo

Andreozzi

Iuliano

et al. 2014

Brain, Behavior, and Immunity

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Structural and functional insights into IκB-α/HIV-1 Tat interaction

Cited by 20 publications

References 53 publications

Immunological and biochemical characterisation of 7ap, a short protein translated from an alternative frame of ORF7 of PRRSV

Immunological and biochemical characterisation of 7ap, a short protein translated from an alternative frame of ORF7 of PRRSV

IBTK Differently Modulates Gene Expression and RNA Splicing in HeLa and K562 Cells

Early molecular and behavioral response to lipopolysaccharide in the WAG/Rij rat model of absence epilepsy and depressive-like behavior, involves interplay between AMPK, AKT/mTOR pathways and neuroinflammatory cytokine release

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