Full-length human cytomegalovirus terminase pUL89 adopts a two-domain structure specific for DNA packaging

English version

Autoři: Janine Theiß ^aff001; Min Woo Sung ^aff002; Andreas Holzenburg ^aff003; Elke Bogner ^aff001
Působiště autorů: Institute of Virology, Charité - Universitätsmedizin Berlin, Berlin, Germany ^aff001; Department of Biochemistry and Molecular Biology, Oregon Health and Science University, Portland, Oregon, United States of America ^aff002; Department of Molecular Science, School of Medicine, The University of Texas Rio Grande Valley, Brownsville-Edinburg-Harlingen, Texas, United States of America ^aff003
Vyšlo v časopise: Full-length human cytomegalovirus terminase pUL89 adopts a two-domain structure specific for DNA packaging. PLoS Pathog 15(12): e32767. doi:10.1371/journal.ppat.1008175
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.ppat.1008175

Souhrn

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.

Klíčová slova:

Bacteriophages – DNA binding assay – DNA-binding proteins – Human cytomegalovirus – Nucleases – Protein structure – Protein structure prediction – Sequence motif analysis

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