Rice, A. P., Carlotti, F. (December 1990) Structural analysis of wild-type and mutant human immunodeficiency virus type 1 Tat proteins. J Virol, 64 (12). pp. 6018-26. ISSN 0022-538X (Print)0022-538X (Linking)
Abstract
We expressed the human immunodeficiency virus type 1 transactivator protein, Tat, in the wheat germ cell-free translation system and found it to exist as a monomer. The first coding exon (residues 1 to 72) of wheat germ-expressed Tat was resistant to trypsin digestion, indicating that it is a highly folded, independently structured protein domain. Several mutant Tat proteins were dramatically more sensitive to trypsin than the wild type was, suggesting that their reduced transactivation activities are the result of destabilized structures. Mutant proteins with single-amino-acid substitutions were also identified that had reduced transactivation activities but wild-type structures in the trypsin assay. These mutants clustered in two regions of Tat, at acidic residues 2 and 5 in the amino terminus and between residues 18 and 32. These mutants, wild type in structure but reduced in activity, identify residues in the wild-type protein that may directly contact other molecules during Tat function.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Alanine Amino Acid Sequence Animals Cell Line Exons Gene Products, tat/*genetics/isolation & purification/metabolism Genes, tat HIV-1/*genetics/metabolism HeLa Cells/metabolism Humans Kinetics Molecular Sequence Data Molecular Weight Mutagenesis, Site-Directed Peptide Mapping Plasmids Protein Biosynthesis Transcriptional Activation Trypsin/metabolism tat Gene Products, Human Immunodeficiency Virus |
| Subjects: | diseases & disorders > viral diseases > HIV bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein characterization |
| CSHL Authors: | |
| Communities: | CSHL labs |
| Depositing User: | Matt Covey |
| Date: | December 1990 |
| Date Deposited: | 09 Feb 2016 15:55 |
| Last Modified: | 09 Feb 2016 15:55 |
| PMCID: | PMC248775 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/32339 |
Actions (login required)
 |
Administrator's edit/view item |