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Full-length human cytomegalovirus terminase pUL89 adopts a two-domain structure specific for DNA packaging
- Source :
- PLoS Pathogens, PLoS Pathogens, Vol 15, Iss 12, p e1008175 (2019)
- Publication Year :
- 2019
- Publisher :
- Public Library of Science, 2019.
-
Abstract
- A key step in replication of human cytomegalovirus (HCMV) in the host cell is the generation and packaging of unit-length genomes into preformed capsids. The enzymes involved in this process are the terminases. The HCMV terminase complex consists of two terminase subunits, the ATPase pUL56 and the nuclease pUL89. A potential third component pUL51 has been proposed. Even though the terminase subunit pUL89 has been shown to be essential for DNA packaging and interaction with pUL56, it is not known how pUL89 mechanistically achieves sequence-specific DNA binding and nicking. To identify essential domains and invariant amino acids vis-a-vis nuclease activity and DNA binding, alanine substitutions of predicted motifs were analyzed. The analyses indicated that aspartate 463 is an invariant amino acid for the nuclease activity, while argine 544 is an invariant aa for DNA binding. Structural analysis of recombinant protein using electron microscopy in conjunction with single particle analysis revealed a curvilinear monomer with two distinct domains connected by a thinner hinge-like region that agrees well with the predicted structure. These results allow us to model how the terminase subunit pUL89’s structure may mediate its function.<br />Author summary HCMV is a member of the herpesvirus family and represents a major human pathogen causing severe disease in newborns and immunocompromised patients for which the development of new non-nucleosidic antiviral agents are highly important. This manuscript focuses on DNA packaging, which is a target for development of new antivirals. The terminase subunit pUL89 is involved in this process. The paper presents the identification of DNA binding and nuclease motifs with invariant amino acids and highlights its first 3-D surface structure at approx. 3 nm resolution. At this resolution, the calculated 3-D surface structure matches well with the predicted structure. In conjunction with earlier studies it was possible to define structure-function relationships for the HCMV terminase subunit pUL89.
- Subjects :
- Hydrolases
Protein Conformation
Cytomegalovirus
Protein Structure Prediction
Pathology and Laboratory Medicine
Biochemistry
law.invention
chemistry.chemical_compound
Database and Informatics Methods
Protein structure
law
Macromolecular Structure Analysis
Medicine and Health Sciences
Bacteriophages
Biology (General)
Alanine
0303 health sciences
biology
Chemistry
030302 biochemistry & molecular biology
Protein structure prediction
Enzymes
Medical Microbiology
Viral Pathogens
Viruses
Recombinant DNA
Human Cytomegalovirus
Chemical characterization
Pathogens
Sequence Analysis
Research Article
Protein Structure
Herpesviruses
QH301-705.5
Nucleases
Bioinformatics
Protein subunit
Immunology
DNA binding assay
DNA-binding protein
Microbiology
03 medical and health sciences
Structure-Activity Relationship
Viral Proteins
Sequence Motif Analysis
Virology
DNA-binding proteins
DNA Packaging
Genetics
Binding analysis
Molecular Biology
Microbial Pathogens
030304 developmental biology
Nuclease
Biology and life sciences
Organisms
Proteins
RC581-607
Research and analysis methods
biology.protein
Biophysics
Enzymology
Parasitology
Immunologic diseases. Allergy
DNA viruses
DNA
Subjects
Details
- Language :
- English
- ISSN :
- 15537374 and 15537366
- Volume :
- 15
- Issue :
- 12
- Database :
- OpenAIRE
- Journal :
- PLoS Pathogens
- Accession number :
- edsair.doi.dedup.....8f4fa79b889d151984c1e14e7a84b674