Mouse cytomegalovirus egress protein pM50 interacts with cellular endophilin-A2

Lemnitzer, Frederic; Raschbichler, Verena; Kolodziejczak, Dominika; Israel, Lars; Imhof, Axel; Bailer, Susanne M.; Koszinowski, Ulrich H.; Ruzsics, Zsolt

doi:10.1111/cmi.12080

Cited by 23 publications

(31 citation statements)

References 85 publications

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“…All herpesviruses analyzed so far utilize a core NEC of pUL50 -pUL53 homologs as a scaffold to assemble further NEC regulators and docking and adaptor proteins (7). However, a comparison between the findings of the present study on human CMV and a related study on murine CMV (54) revealed that the findings were only partly consistent. The central NEC components pUL50/pM50, pUL53/pM53, and p32/gC1qR were identified accordingly, but a huge number of NEC-associated proteins differed between the two systems.…”

Section: Differential Nec Assembly Comparing Human and Murinecontrasting

confidence: 55%

Proteomic Analysis of the Multimeric Nuclear Egress Complex of Human Cytomegalovirus

Milbradt

Kraut

Hutterer

et al. 2014

Molecular & Cellular Proteomics

110

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Herpesviral capsids are assembled in the host cell nucleus before being translocated into the cytoplasm for further maturation. The crossing of the nuclear envelope represents a major event that requires the formation of the nuclear egress complex (NEC). Previous studies demonstrated that human cytomegalovirus (HCMV) proteins pUL50 and pUL53, as well as their homologs in all members of Herpesviridae, interact with each other at the nuclear envelope and form the heterodimeric core of the NEC. In order to characterize further the viral and cellular protein content of the multimeric NEC, the native complex was isolated from HCMV-infected human primary fibroblasts at various time points and analyzed using quantitative proteomics. Previously postulated components of the HCMVspecific NEC, as well as novel potential NEC-associated proteins such as emerin, were identified. In this regard, interaction and colocalization between emerin and pUL50 were confirmed by coimmunoprecipitation and confocal microscopy analyses, respectively. A functional validation of viral and cellular NEC constituents was achieved through siRNA-mediated knockdown experiments. The important role of emerin in NEC functionality was demonstrated by a reduction of viral replication when emerin expression was down-regulated. Moreover, under such conditions, reduced production of viral proteins and deregulation of viral late cytoplasmic maturation were observed. Combined, these data prove the functional importance of emerin as an NEC component, associated with pUL50, pUL53, pUL97, p32/ gC1qR, and further regulatory proteins. Summarized, our findings provide the first proteomics-based characterization and functional validation of the HCMV-specific multimeric NEC. Molecular & Cellular

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Section: Differential Nec Assembly Comparing Human and Murinecontrasting

confidence: 55%

Proteomic Analysis of the Multimeric Nuclear Egress Complex of Human Cytomegalovirus

Milbradt

Kraut

Hutterer

et al. 2014

Molecular & Cellular Proteomics

110

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“…Various lines of experimental evidence suggested that the cytomegaloviral core NEC acts as a scaffold to assemble accessory NEC components (Muranyi et al, 2002;Milbradt et al, 2007Milbradt et al, , 2009Milbradt et al, , 2014Buchkovich et al, 2010;Lemnitzer et al, 2013). In particular, pUL50 defines the formation of the multimeric NEC by interacting with pUL53, PKCa, p32/gC1qR and emerin.…”

Section: Core Nec Formation Is Sensitive To Inhibitors Of Cdk Activitymentioning

confidence: 99%

Cytomegalovirus pUL50 is the multi-interacting determinant of the core nuclear egress complex (NEC) that recruits cellular accessory NEC components

Sonntag

Hamilton

Hanife

et al. 2016

Journal of General Virology

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Nuclear egress of herpesvirus capsids through the nuclear envelope is mediated by the multimeric nuclear egress complex (NEC). The human cytomegalovirus (HCMV) core NEC is defined by an interaction between the membrane-anchored pUL50 and its nuclear co-factor pUL53, tightly associated through heterodimeric corecruitment to the nuclear envelope. Cellular proteins, such as p32/gC1qR, emerin and protein kinase C (PKC), are recruited by direct interaction with pUL50 for the multimeric extension of the NEC. As a functionally important event, the recruitment of both viral and cellular protein kinases leads to site-specific lamin phosphorylation and nuclear lamina disassembly. In this study, interaction domains within pUL50 for its binding partners were defined by co-immunoprecipitation. The interaction domain for pUL53 is located within the pUL50 N-terminus (residues 10-169), interaction domains for p32/gC1qR (100-358) and PKC (100-280) overlap in the central part of pUL50, and the interaction domain for emerin is located in the C-terminus (265-397). Moreover, expression and formation of core NEC proteins at the nuclear rim were consistently detected in cells permissive for productive HCMV replication, including two trophoblast-cell lines. Importantly, regular nuclear-rim formation of the core NEC was blocked by inhibition of cyclin-dependent kinase (CDK) activity. In relation to the recently published crystal structure of the HCMV core NEC, our findings result in a refined view of NEC assembly. In particular, we suggest that CDKs may play an important regulatory role in NEC formation during HCMV replication.

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“…Recently published proteomic data provided for the first time a candidate list of the associated viral and cellular constituents of the multimeric NEC for both HCMV and MCMV (15,18). For HCMV, at least four proteins were identified that directly interact with pUL50, i.e.…”

Section: Resultsmentioning

confidence: 99%

“…As an essential step in HCMV replication, pUL50 and pUL53 heterodimerize and form the core of the NEC that serves as a scaffold for the recruitment of a group of viral and cellular NEC-associated proteins. The composition of the multimeric NEC of human and murine CMVs has recently been defined by proteomic analyses (15,18).…”

mentioning

confidence: 99%

Crystal Structure of the Human Cytomegalovirus pUL50-pUL53 Core Nuclear Egress Complex Provides Insight into a Unique Assembly Scaffold for Virus-Host Protein Interactions

Walzer

Egerer-Sieber

Sticht

et al. 2015

Journal of Biological Chemistry

139

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Background:The conserved cytomegalovirus proteins pUL50 and pUL53 heterodimerize and form a core nuclear egress complex. Results: The crystal structure of pUL50-pUL53 was solved at 2.44 Å resolution, revealing an N-terminal hooklike extension of pUL53. Conclusion: Data unravel the core NEC architecture, providing a scaffold for viral-cellular NEC protein interactions. Significance: The identified NEC structure will stimulate the development of novel antiviral strategies.

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Mouse cytomegalovirus egress protein pM50 interacts with cellular endophilin-A2

Cited by 23 publications

References 85 publications

Proteomic Analysis of the Multimeric Nuclear Egress Complex of Human Cytomegalovirus

Proteomic Analysis of the Multimeric Nuclear Egress Complex of Human Cytomegalovirus

Cytomegalovirus pUL50 is the multi-interacting determinant of the core nuclear egress complex (NEC) that recruits cellular accessory NEC components

Crystal Structure of the Human Cytomegalovirus pUL50-pUL53 Core Nuclear Egress Complex Provides Insight into a Unique Assembly Scaffold for Virus-Host Protein Interactions

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