The ICP27 Homology Domain of the Human Cytomegalovirus Protein UL69 Adopts a Dimer-of-Dimers Structure

ABSTRACT The UL69 protein from human cytomegalovirus (HCMV) is a multifunctional regulatory protein and a member of the ICP27 protein family conserved throughout herpesviruses. UL69 plays many roles during productive infection, including the regulation of viral gene expression, nuclear export of int...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Richard B. Tunnicliffe, Richard F. Collins, Hilda D. Ruiz Nivia, Rozanne M. Sandri-Goldin, Alexander P. Golovanov
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	human cytomegalovirus ICP27 homology domain structural analysis tetramerization transmission electron microscopy Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/59fa04a094154df3b18d193debc92a9a
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:59fa04a094154df3b18d193debc92a9a
record_format	dspace
spelling	oai:doaj.org-article:59fa04a094154df3b18d193debc92a9a2021-11-15T16:00:27ZThe ICP27 Homology Domain of the Human Cytomegalovirus Protein UL69 Adopts a Dimer-of-Dimers Structure10.1128/mBio.01112-182150-7511https://doaj.org/article/59fa04a094154df3b18d193debc92a9a2018-07-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01112-18https://doaj.org/toc/2150-7511ABSTRACT The UL69 protein from human cytomegalovirus (HCMV) is a multifunctional regulatory protein and a member of the ICP27 protein family conserved throughout herpesviruses. UL69 plays many roles during productive infection, including the regulation of viral gene expression, nuclear export of intronless viral RNAs, and control of host cell cycle progression. Throughout the ICP27 protein family, an ability to self-associate is correlated with the functions of these proteins in transactivating certain viral genes. Here, we determined the domain boundaries of a globular ICP27 homology domain of UL69, which mediates self-association, and characterized the oligomeric state of the isolated domain. Size exclusion chromatography coupled with multiangle light scattering (SEC-MALS) revealed that residues 200 to 540 form a stable homo-tetramer, whereas a shorter region comprising residues 248 to 536 forms a homo-dimer. Structural analysis of the UL69 tetramer by transmission electron microscopy (TEM) revealed a dimer-of-dimers three-dimensional envelope with bridge features likely from a region of the protein unique to betaherpesviruses. The data provide a structural template for tetramerization and improve our understanding of the structural diversity and features necessary for self-association within UL69 and the ICP27 family. IMPORTANCE Human cytomegalovirus (HCMV) infection is widespread in the human population but typically remains dormant in an asymptomatic latent state. HCMV causes disease in neonates and adults with suppressed or impaired immune function, as the virus is activated into a lytic state. All species of herpesvirus express a protein from the ICP27 family which functions as a posttranscriptional activator in the lytic state. In HCMV, this protein is called UL69. The region of sequence conservation in the ICP27 family is a folded domain that mediates protein interactions, including self-association and functions in transactivation. All members thus far analyzed homo-dimerize, with the exception of UL69, which forms higher-order oligomers. Here, we use biochemical and structural data to reveal that UL69 forms stable tetramers composed of a dimer of dimers and determine a region essential for cross-dimer stabilization.Richard B. TunnicliffeRichard F. CollinsHilda D. Ruiz NiviaRozanne M. Sandri-GoldinAlexander P. GolovanovAmerican Society for Microbiologyarticlehuman cytomegalovirusICP27 homology domainstructural analysistetramerizationtransmission electron microscopyMicrobiologyQR1-502ENmBio, Vol 9, Iss 3 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	human cytomegalovirus ICP27 homology domain structural analysis tetramerization transmission electron microscopy Microbiology QR1-502
spellingShingle	human cytomegalovirus ICP27 homology domain structural analysis tetramerization transmission electron microscopy Microbiology QR1-502 Richard B. Tunnicliffe Richard F. Collins Hilda D. Ruiz Nivia Rozanne M. Sandri-Goldin Alexander P. Golovanov The ICP27 Homology Domain of the Human Cytomegalovirus Protein UL69 Adopts a Dimer-of-Dimers Structure
description	ABSTRACT The UL69 protein from human cytomegalovirus (HCMV) is a multifunctional regulatory protein and a member of the ICP27 protein family conserved throughout herpesviruses. UL69 plays many roles during productive infection, including the regulation of viral gene expression, nuclear export of intronless viral RNAs, and control of host cell cycle progression. Throughout the ICP27 protein family, an ability to self-associate is correlated with the functions of these proteins in transactivating certain viral genes. Here, we determined the domain boundaries of a globular ICP27 homology domain of UL69, which mediates self-association, and characterized the oligomeric state of the isolated domain. Size exclusion chromatography coupled with multiangle light scattering (SEC-MALS) revealed that residues 200 to 540 form a stable homo-tetramer, whereas a shorter region comprising residues 248 to 536 forms a homo-dimer. Structural analysis of the UL69 tetramer by transmission electron microscopy (TEM) revealed a dimer-of-dimers three-dimensional envelope with bridge features likely from a region of the protein unique to betaherpesviruses. The data provide a structural template for tetramerization and improve our understanding of the structural diversity and features necessary for self-association within UL69 and the ICP27 family. IMPORTANCE Human cytomegalovirus (HCMV) infection is widespread in the human population but typically remains dormant in an asymptomatic latent state. HCMV causes disease in neonates and adults with suppressed or impaired immune function, as the virus is activated into a lytic state. All species of herpesvirus express a protein from the ICP27 family which functions as a posttranscriptional activator in the lytic state. In HCMV, this protein is called UL69. The region of sequence conservation in the ICP27 family is a folded domain that mediates protein interactions, including self-association and functions in transactivation. All members thus far analyzed homo-dimerize, with the exception of UL69, which forms higher-order oligomers. Here, we use biochemical and structural data to reveal that UL69 forms stable tetramers composed of a dimer of dimers and determine a region essential for cross-dimer stabilization.
format	article
author	Richard B. Tunnicliffe Richard F. Collins Hilda D. Ruiz Nivia Rozanne M. Sandri-Goldin Alexander P. Golovanov
author_facet	Richard B. Tunnicliffe Richard F. Collins Hilda D. Ruiz Nivia Rozanne M. Sandri-Goldin Alexander P. Golovanov
author_sort	Richard B. Tunnicliffe
title	The ICP27 Homology Domain of the Human Cytomegalovirus Protein UL69 Adopts a Dimer-of-Dimers Structure
title_short	The ICP27 Homology Domain of the Human Cytomegalovirus Protein UL69 Adopts a Dimer-of-Dimers Structure
title_full	The ICP27 Homology Domain of the Human Cytomegalovirus Protein UL69 Adopts a Dimer-of-Dimers Structure
title_fullStr	The ICP27 Homology Domain of the Human Cytomegalovirus Protein UL69 Adopts a Dimer-of-Dimers Structure
title_full_unstemmed	The ICP27 Homology Domain of the Human Cytomegalovirus Protein UL69 Adopts a Dimer-of-Dimers Structure
title_sort	icp27 homology domain of the human cytomegalovirus protein ul69 adopts a dimer-of-dimers structure
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/59fa04a094154df3b18d193debc92a9a
work_keys_str_mv	AT richardbtunnicliffe theicp27homologydomainofthehumancytomegalovirusproteinul69adoptsadimerofdimersstructure AT richardfcollins theicp27homologydomainofthehumancytomegalovirusproteinul69adoptsadimerofdimersstructure AT hildadruiznivia theicp27homologydomainofthehumancytomegalovirusproteinul69adoptsadimerofdimersstructure AT rozannemsandrigoldin theicp27homologydomainofthehumancytomegalovirusproteinul69adoptsadimerofdimersstructure AT alexanderpgolovanov theicp27homologydomainofthehumancytomegalovirusproteinul69adoptsadimerofdimersstructure AT richardbtunnicliffe icp27homologydomainofthehumancytomegalovirusproteinul69adoptsadimerofdimersstructure AT richardfcollins icp27homologydomainofthehumancytomegalovirusproteinul69adoptsadimerofdimersstructure AT hildadruiznivia icp27homologydomainofthehumancytomegalovirusproteinul69adoptsadimerofdimersstructure AT rozannemsandrigoldin icp27homologydomainofthehumancytomegalovirusproteinul69adoptsadimerofdimersstructure AT alexanderpgolovanov icp27homologydomainofthehumancytomegalovirusproteinul69adoptsadimerofdimersstructure
_version_	1718426967907237888

The ICP27 Homology Domain of the Human Cytomegalovirus Protein UL69 Adopts a Dimer-of-Dimers Structure

Ejemplares similares