Recovery of an HMWP/hmwBP (pUL48/pUL47) complex from virions of human cytomegalovirus: Subunit interactions, oligomer composition, and deubiquitylase activity

Jennifer A. Tullman, Mary Elizabeth Harmon, Michael Delannoy, D Wade Gibson

Research output: Contribution to journalArticle

Abstract

We report that the human cytomegalovirus (HCMV) high-molecular-weight tegument protein (HMWP, pUL48; 253 kDa) and the HMWP-binding protein (hmwBP, pUL47; 110 kDa) can be recovered as a complex from virions disrupted by treatment with 50 mM Tris (pH 7.5), 0.5 M NaCl, 0.5% NP-40, and 10 mM dithiothreitol [DTT]. The subunit ratio of the complex approximates 1:1, with a shape and structure consistent with an elongated heterodimer. The HMWP/hmwBP complex was corroborated by reciprocal coimmunoprecipitation experiments using antipeptide antibodies and lysates from both infected cells and disrupted virus particles. An interaction of the amino end of pUL48 (amino acids [aa] 322 to 754) with the carboxyl end of pUL47 (aa 693 to 982) was identified by fragment coimmunoprecipitation experiments, and a head-to-tail self-interaction of hmwBP was also observed. The deubiquitylating activity of pUL48 is retained in the isolated complex, which cleaves K11, K48, and K63 ubiquitin isopeptide linkages.

Original languageEnglish (US)
Pages (from-to)8256-8267
Number of pages12
JournalJournal of Virology
Volume88
Issue number15
DOIs
StatePublished - 2014

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Human herpesvirus 5
Cytomegalovirus
virion
Virion
Amino Acids
amino acids
dithiothreitol
Dithiothreitol
ubiquitin
Ubiquitin
binding proteins
Carrier Proteins
Molecular Weight
molecular weight
antibodies
Antibodies
Proteins
proteins
cells
Nonidet P-40

ASJC Scopus subject areas

  • Immunology
  • Virology

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Recovery of an HMWP/hmwBP (pUL48/pUL47) complex from virions of human cytomegalovirus : Subunit interactions, oligomer composition, and deubiquitylase activity. / Tullman, Jennifer A.; Harmon, Mary Elizabeth; Delannoy, Michael; Gibson, D Wade.

In: Journal of Virology, Vol. 88, No. 15, 2014, p. 8256-8267.

Research output: Contribution to journalArticle

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