Characterization of the Acidic Domain of the IE1 Regulatory Protein fromOrgyia pseudotsugataMulticapsid Nucleopolyhedrovirus
| Autor: | Cynthia E. Shippam, Leslie G. Willis, Ian J. Forsythe, David A. Theilmann, T. A. Grigliatti, Sandra Stewart |
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| Rok vydání: | 1998 |
| Předmět: |
viruses
Recombinant Fusion Proteins Molecular Sequence Data Sequence alignment Biology Spodoptera Cell Line Immediate-Early Proteins 03 medical and health sciences Transactivation Virology Animals Amino Acid Sequence Peptide sequence 030304 developmental biology chemistry.chemical_classification 0303 health sciences Reporter gene fungi 030302 biochemistry & molecular biology Herpes Simplex Virus Protein Vmw65 biochemical phenomena metabolism and nutrition biology.organism_classification Molecular biology Nucleopolyhedroviruses Amino acid DNA-Binding Proteins Autographa californica Biochemistry chemistry Trans-Activators Orgyia pseudotsugata Sequence Alignment |
| Zdroj: | Virology. 252(1):65-81 |
| ISSN: | 0042-6822 |
| DOI: | 10.1006/viro.1998.9415 |
| Popis: | This study presents a detailed analysis of the acidic N-terminal region of theOrgyia pseudotsugatamulticapsid nucleopolyhedrovirus (OpMNPV) transactivator IE1. The N-terminal region of IE1 is rich in acidic amino acids and has been hypothesized to be an acidic activation domain. Removal of the N-terminal 126 amino acids containing the acidic domain of IE1 resulted in complete loss of transactivation activity, indicating that this region is essential for transactivation. The OpMNPV acidic domain was replaced with the archetype acidic activation domain from VP16 and the acid-rich region ofAutographa californicamulticapsid NPV (AcMNPV) IE1. These chimeric constructs were fully capable of transactivation in transient assays. The chimeric OpMNPV IE1s containing the herpes simplex virus VP16 and AcMNPV IE1 acidic activation domains consistently transactivated a reporter gene to higher levels than the OpMNPV IE1 acidic activation domain. Transactivation by the chimeric constructs is enhanced synergistically when cotransfected with IE2 intoLymantria disparandSpodoptera frugiperdacells. Both N- to C-terminal and C- to N-terminal deletions of the OpMNPV acidic activation domain were constructed to define functional domains within the OpMNPV IE1 acidic activation domain. At least two potential activation domains were identified. Within each of these domains, two core regions at amino acids 28–43 and amino acids 113–124 were identified that were similar to core regions of VP16 and GAL4, which contain predominately acidic and bulky hydrophobic amino acids. |
| Databáze: | OpenAIRE |
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