Protein Array Identification of Substrates of the Epstein-Barr Virus Protein Kinase BGLF4
- 15 May 2009
- journal article
- Published by American Society for Microbiology in Journal of Virology
- Vol. 83 (10), 5219-5231
- https://doi.org/10.1128/jvi.02378-08
Abstract
A conserved family of herpesvirus protein kinases plays a crucial role in herpesvirus DNA replication and virion production. However, despite the fact that these kinases are potential therapeutic targets, no systematic studies have been performed to identify their substrates. We generated an Epstein-Barr virus (EBV) protein array to evaluate the targets of the EBV protein kinase BGLF4. Multiple proteins involved in EBV lytic DNA replication and virion assembly were identified as previously unrecognized substrates for BGLF4, illustrating the broad role played by this protein kinase. Approximately half of the BGLF4 targets were also in vitro substrates for the cellular kinase CDK1/cyclin B. Unexpectedly, EBNA1 was identified as a substrate and binding partner of BGLF4. EBNA1 is essential for replication and maintenance of the episomal EBV genome during latency. BGLF4 did not prevent EBNA1 binding to sites in the EBV latency origin of replication, oriP . Rather, we found that BGLF4 was recruited by EBNA1 to oriP in cells transfected with an oriP vector and BGLF4 and in lytically induced EBV-positive Akata cells. In cells transfected with an oriP vector, the presence of BGLF4 led to more rapid loss of the episomal DNA, and this was dependent on BGLF4 kinase activity. Similarly, expression of doxycycline-inducible BGLF4 in Akata cells led to a reduction in episomal EBV genomes. We propose that BGLF4 contributes to effective EBV lytic cycle progression, not only through phosphorylation of EBV lytic DNA replication and virion proteins, but also by interfering with the EBNA1 replication function.Keywords
This publication has 46 references indexed in Scilit:
- Functional Dissection of a HECT Ubiquitin E3 LigaseMolecular & Cellular Proteomics, 2008
- Conserved herpesvirus protein kinasesBiochimica et Biophysica Acta (BBA) - Proteins and Proteomics, 2008
- A proteome chip approach reveals new DNA damage recognition activities in Escherichia coliNature Methods, 2007
- γ-Herpesvirus Kinase Actively Initiates a DNA Damage Response by Inducing Phosphorylation of H2AX to Foster Viral ReplicationCell Host & Microbe, 2007
- Epstein-Barr Virus BGLF4 Kinase Induces Premature Chromosome Condensation through Activation of Condensin and Topoisomerase IIJournal of Virology, 2007
- Epstein–Barr virus and virus human protein interaction mapsProceedings of the National Academy of Sciences of the United States of America, 2007
- Ubiquitination screen using protein microarrays for comprehensive identification of Rsp5 substrates in yeastMolecular Systems Biology, 2007
- Phosphorylation of MCM4 at Sites Inactivating DNA Helicase Activity of the MCM4-MCM6-MCM7 Complex during Epstein-Barr Virus Productive ReplicationJournal of Virology, 2006
- The infectious kiss: Newly infected B cells deliver Epstein–Barr virus to epithelial cellsProceedings of the National Academy of Sciences of the United States of America, 2006
- Epstein-Barr virus: exploiting the immune systemNature Reviews Immunology, 2001