ArticleProceedings of the National Academy of Sciences of the United States of America1989
Bioassay for trans-activation using purified human immunodeficiency virus tat-encoded protein: trans-activation requires mRNA synthesis.
Article in Proceedings of the National Academy of Sciences of the United States of America, 1989. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 48 papers.
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Who cites it
48 citing papers in PubMed, 123 citations in OpenAlex.
- Intracellular Delivery by Membrane Disruption: Mechanisms, Strategies, and Concepts.Chemical reviews · 2018Review
- Chemically well-defined self-assembled monolayers for cell culture: toward mimicking the natural ECM.Soft matter · 2011Article
- Cell-to-cell spread of the RNA interference response suppresses Semliki Forest virus (SFV) infection of mosquito cell cultures and cannot be antagonized by SFV.Journal of virology · 2009Article
- Interpretation of matrix-assisted laser desorption/ionization postsource decay spectra of charge-derivatized peptides: some examples of tris[(2,4,6-trimethoxyphenyl) phosphonium]-tagged proteolytic digestion products of phosphoenolpyruvate carboxykinase.Journal of the American Society for Mass Spectrometry · 2000Article
- Tat protein induces human immunodeficiency virus type 1 (HIV-1) coreceptors and promotes infection with both macrophage-tropic and T-lymphotropic HIV-1 strains.Journal of virology · 1998Article
- Modulation of Sp1 phosphorylation by human immunodeficiency virus type 1 Tat.Journal of virology · 1998Article
- Characterization of two mutations associated with epimerase-deficiency galactosemia, by use of a yeast expression system for human UDP-galactose-4-epimerase.American journal of human genetics · 1997Article
- Tat protein induces self-perpetuating permissivity for productive HIV-1 infection.Proceedings of the National Academy of Sciences of the United States of America · 1997Article
- Identification of the coding region for a second poly(A) polymerase in Escherichia coli.Proceedings of the National Academy of Sciences of the United States of America · 1996Article
- Heterodimer formation and activity in the human enzyme galactose-1-phosphate uridylyltransferase.Proceedings of the National Academy of Sciences of the United States of America · 1996Article
- The catalytic core of RNase P.Nucleic acids research · 1996Article
- One-step purification of recombinant proteins with the 6xHis tag and Ni-NTA resin.Molecular biotechnology · 1995Article
- Wilms' tumor 1 susceptibility (WT1) gene products are selectively expressed in malignant mesothelioma.The American journal of pathology · 1995Article
- HIV-1 Tat potentiates TNF-induced NF-kappa B activation and cytotoxicity by altering the cellular redox state.The EMBO journal · 1995Article
- Extracellular Vpr protein increases cellular permissiveness to human immunodeficiency virus replication and reactivates virus from latency.Journal of virology · 1995Article
- Human immunodeficiency virus 1 Tat binds to dipeptidyl aminopeptidase IV (CD26): a possible mechanism for Tat's immunosuppressive activity.Proceedings of the National Academy of Sciences of the United States of America · 1994Article
- Characterization and cloning of the human splicing factor 9G8: a novel 35 kDa factor of the serine/arginine protein family.The EMBO journal · 1994Article
- The relationship between serum p53 autoantibodies and characteristics of human breast cancer.British journal of cancer · 1994Article
- SRY protein enhances transcription of Fos-related antigen 1 promoter constructs.Proceedings of the National Academy of Sciences of the United States of America · 1994Article
- Inhibition of human immunodeficiency virus type 1 Tat-dependent activation of translation in Xenopus oocytes by the benzodiazepine Ro24-7429 requires trans-activation response element loop sequences.Journal of virology · 1994Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Expression of the human immunodeficiency virus tat-encoded protein (Tat) is required for virus replication. A genetic approach was used to facilitate the purification of biologically active Tat. A recombinant Tat protein containing a stretch of six histidine residues and a protease cleavage site was engineered and purified to greater than 95% homogeneity in a single step by immobilized metal-ion chromatography with a special affinity resin that has selectivity for proteins with neighboring histidine residues. A modified scrape loading method for introduction of protein into cell monolayers was used to demonstrate that the purified Tat retained biological activity. Tat function was completely blocked in the presence of transcription inhibitors, which demonstrates the requirement of ongoing mRNA synthesis for trans-activation. These studies indicate that the mechanism of trans-activation is unlikely to involve a direct action of Tat on mRNA stability, transport, or translation and provides the basis for a rapid assay that can be used to identify inhibitors of trans-activation. The methods described herein should be useful for the functional analysis of other proteins that do not confer activity through a receptor-mediated pathway.
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