Herpes simplex Virus Type 1 DNase: Functional Analysis of the Enzyme Expressed by Recombinant Baculovirus
| Autor: | Mehmet-Ali Göksu, Stefanie Bayer, Charles W. Knopf, Elke Kehm |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Genes
Viral viruses Gene Expression Herpesvirus 1 Human Spodoptera Virus Replication medicine.disease_cause Alkaline DNase Cell Line law.invention Endonuclease law Cricetinae Virology medicine Animals Cloning Molecular Recombination Genetic chemistry.chemical_classification Deoxyribonucleases biology biology.organism_classification Molecular biology Nucleopolyhedroviruses Recombinant Proteins Molecular Weight Infectious Diseases Herpes simplex virus Enzyme chemistry Viral replication Cell culture DNA Viral Recombinant DNA biology.protein Dimerization |
| Zdroj: | Intervirology. 41:110-119 |
| ISSN: | 1423-0100 0300-5526 |
| Popis: | Herpes simplex virus type 1 DNase (HSV-1 DNase) was expressed in insect cells by recombinant baculovirus (NPVUL12) and purified by a combination of anionic exchanger chromatography and gel filtration. Two polypeptides of 85 and 75 kD, whose ratio varied during purification, were induced 24 h after infection. The 75-kD protein was isolated and shown to possess catalytic activity. Gel filtration analysis indicated that the active form of the enzyme at an ionic strength of I = 0.3 is a dimeric protein with an apparent molecular weight of 130,000. The recombinant enzyme exhibited the overall characteristics of the native enzyme such as 5′-3′ exonuclease and endonuclease activities with a preferred degradation of DNA. In the absence of extraneously added Mg2+, the enzyme was capable of removing mononucleotides from 5′-end-labeled DNA, but not from RNA and 3′-end-labeled DNA. The peculiar mechanism of double-strand DNA degradation suggests a specific role of HSV-1 DNase in DNA recombination processes during viral replication. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|