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Analysis of the interactions of viral and cellular factors with human cytomegalovirus lytic origin of replication, oriLyt
Open accessAbstract
Human cytomegalovirus transient lytic DNA replication relies on the cis-acting element oriLyt, six viral-encoded core proteins, the proposed DNA replication initiator protein UL84, IE2, IRS1 and the gene products from the UL112/113 loci. In an effort to elucidate cellular and viral-encoded factors that may play a role in oriLyt-dependent replication we used DNA-affinity purification and mass spectrometry to isolate and identify several previously unknown cellular and viral factors that interact with HCMV oriLyt DNA. These proteins include the multifunctional hnRNP-K, BUB3, HMGB1, PTB-1, UL83, UL112/113, and IRS1. Chromatin immunoprecipitation (ChIP) assays confirmed an interaction of several of these factors with oriLyt. Co-immunoprecipitation experiments detected an interaction between UL84 and hnRNP-K in infected and transfected cells. Knockdown of hnRNP K expression by siRNA inhibited the amplification of oriLyt in the transient assay. Together, these data suggest a possible regulatory role in DNA replication for several previously unidentified viral and cellular factors.
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“AP-MS has been employed to determine the identity of such host proteins, for instance, proteins in replication complexes at viral promoters in herpesviruses (67–69), viral polymerases of influenza virus (53), and preintegration complexes in HIV-1 (70).”
From The Impact of Mass Spectrometry–Based Proteomics on Fundamental Discoveries in Virology (2014) · cites this paper as [68]
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References 28 · Cited by 32
Research works printed but not linked: 1
- Pari, 1993, Eleven loci encoding trans-acting factors are required for transient complementation of human cytomegalovirus oriLyt-dependent DNA replication, J. Virol., 67, 6979, 10.1128/jvi.67.12.6979-6988.1993 · DOI
Metadata
- Authors
- Kagele, D., Rossetto, C. C., Tarrant, M. T., & Pari, G. S.
- Affiliations
- University of Nevada, Reno (US)
- Corresponding author
- Pari, Gregory S.
- ORCID
- Rossetto, Cyprian C.
- Venue
- Virology
- Publisher
- Elsevier BV
- Published
- 2012-03
- Volume
- 424
- Issue
- 2
- Pages
- 106-114
- ISSN
- 0042-6822 (ISSN-L)
- PMC
- PMC3288578
- OpenAlex
- W2061570282
- Cited by
- 32
- References
- 28
- Citations by year
- 2026: 1 · 2024: 3 · 2023: 1 · 2022: 5 · 2021: 3 · 2020: 1 · 2019: 1 · 2018: 1 · 2017: 1 · 2015: 1 · 2014: 6 · 2013: 5 · 2012: 3
- FWCI
- 3.59
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Virology (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- NIH Public health service grant (R01AI045096)
- Topics
- Health Sciences › Medicine › Epidemiology › Cytomegalovirus and herpesvirus research
- Health Sciences › Medicine › Oncology › Viral-associated cancers and disorders
- Health Sciences › Medicine › Infectious Diseases › Herpesvirus Infections and Treatments
- Keywords
- human cytomegalovirus, origin of replication, hnRNP K, IRS-1, IE2, Bub3, HMGB1, protein-DNA interactions
- MeSH terms
- Poly-ADP-Ribose Binding Proteins, Cell Line, Cytomegalovirus Infections, Cytomegalovirus, DNA Replication, Humans, Protein Binding, Viral Proteins, Virus Replication, Replication Origin, Cell Cycle Proteins, Polypyrimidine Tract-Binding Protein, Host-Pathogen Interactions
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