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Published on: December 21, 2019
Duck plague virus pUL15 performs a nonspecial cleavage activity through its C terminal nuclease domain in vitro
Qiao Yang1, Yiheng Liu1, Mingshu Wang1
1Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu City, Sichuan 611130, China; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Wenjiang, Chengdu City, Sichuan 611130, China; Avian Disease Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu City, Sichuan 611130, China.
Abstract:
Duck plague virus (DPV), also known as anatid herpesvirus, is a double-stranded DNA virus and a member of α herpesvirus. DPV pUL15 is a homolog of herpes simplex virus 1 (HSV-1) pUL15, a terminase large subunit, and plays a key role in the cleavage and packaging of the viral concatemeric genome. However, the sequence similarity between DPV pUL15 and its homologs is low, and it is not sure if DPV pUL15 has the potential to cleave the concatemeric genome as same as its homologs. Here, we expressed the C terminal domain of DPV pUL15 to explore the nuclease function of DPV pUL15. The main results showed that (Ⅰ) DPV pUL15 C-terminal domain possesses nonspecific nuclease activity and lacks the DNA binding ability. (Ⅱ) DPV pUL15 nuclease activity needs to coordinate with divalent metal ions and tends to be more active at high temperatures. (Ⅲ) Even though the structure of DPV pUL15 nuclease domain is relatively conserved, the mutations of conserved amino acids on the nuclease domain do not significantly inhibit the nuclease activity.
Insights
Duck plague virus pUL15 exhibits nonspecific nuclease activity, independent of DNA binding. This activity requires metal ions and functions optimally at high temperatures, even with mutations in conserved regions.
Area of Science:
- Virology
- Molecular Biology
- Biochemistry
Background:
- Duck plague virus (DPV), an anatid herpesvirus, is an alpha herpesvirus.
- DPV pUL15 is a homolog of HSV-1 pUL15, crucial for viral genome packaging.
- Low sequence similarity of DPV pUL15 to homologs raises questions about its function.
Purpose of the Study:
- To investigate the nuclease function of the C-terminal domain of DPV pUL15.
- To determine if DPV pUL15 can cleave the viral concatemeric genome.
Main Methods:
- Expression of the C-terminal domain of DPV pUL15.
- Assays to evaluate nuclease activity and DNA binding.
- Site-directed mutagenesis of conserved amino acids.
Main Results:
- DPV pUL15 C-terminal domain shows nonspecific nuclease activity and lacks DNA binding.
- Nuclease activity is dependent on divalent metal ions and optimal at high temperatures.
- Mutations in conserved nuclease domain residues do not significantly impair activity.
Conclusions:
- DPV pUL15 possesses intrinsic, albeit nonspecific, nuclease activity.
- The enzyme's function is modulated by metal ions and temperature.
- Conserved residues are not essential for the observed nuclease function, suggesting unique mechanisms.
