biostudies-literatureMiller JTHHS | National Institutes of HealthCCR NIH HHSHHS | NIH | National Institute of General Medical SciencesHHS | NIH | National Cancer InstituteNCI NIH HHSNIGMS NIH HHSe00233-18https://www.ebi.ac.uk/biostudies/studies/S-EPMC6153795biostudies-literatureUnknown62(10)Kaposi's sarcoma-associated herpesvirus (KSHV), the etiological agent of Kaposi's sarcoma, belongs to the <i>Herpesviridae</i> family, whose members employ a multicomponent terminase to resolve nonparametric viral DNA into genome-length units prior to their packaging. Homology modeling of the ORF29 C-terminal nuclease domain (pORF29C) and bacteriophage Sf6 gp2 have suggested an active site clustered with four acidic residues, D⁴⁷⁶, E⁵⁵⁰, D⁶⁶¹, and D⁶⁶², that collectively sequester the catalytic divalent metal (Mn²⁺) and also provided important insight into a potential inhibitor binding mode. Using this model, we have expressed, purified, and characterized the wild-type pORF29C and variants with substitutions at the proposed active-siteAntimicrobial agents and chemotherapySensitivity of the C-Terminal Nuclease Domain of Kaposi's Sarcoma-Associated Herpesvirus ORF29 to Two Classes of Active-Site Ligands.PMC6153795R01 GM090010SC1 GM111158HHSN261200800001ER01GM090010HHSN261200800001CSC1GM111158Miller JTAron KEHirsch DRKouhestani KCornejo Castro EMMarshall VATang LLe Grice SFJMurelli RPVarnado BXia ABeutler JAWhitby DZhao HMasaoka TLynn AYfalseSensitivity of the C-Terminal Nuclease Domain of Kaposi's Sarcoma-Associated Herpesvirus ORF29 to Two Classes of Active-Site Ligands.Kaposi's sarcoma-associated herpesvirus (KSHV), the etiological agent of Kaposi's sarcoma, belongs to the <i>Herpesviridae</i> family, whose members employ a multicomponent terminase to resolve nonparametric viral DNA into genome-length units prior to their packaging. Homology modeling of the ORF29 C-terminal nuclease domain (pORF29C) and bacteriophage Sf6 gp2 have suggested an active site clustered with four acidic residues, D⁴⁷⁶, E⁵⁵⁰, D⁶⁶¹, and D⁶⁶², that collectively sequester the catalytic divalent metal (Mn²⁺) and also provided important insight into a potential inhibitor binding mode. Using this model, we have expressed, purified, and characterized the wild-type pORF29C and variants with substitutions at the proposed active-site2018-01-01T00:00:00Z2018 Oct2026-06-14T04:19:55.78Z2026-06-14T03:09:08.253ZS-EPMC61537953006127810.1128/aac.00233-1810.1128/AAC.00233-18