title: Association of Human Papillomavirus 16 E2 with Rad50-Interacting Protein 1 Enhances Viral DNA Replication.
abstract: Rad50-interacting protein 1 (Rint1) associates with the DNA damage response protein Rad50 during the transition from the S phase to the G2/M phase and functions in radiation-induced G2checkpoint control. It has also been demonstrated that Rint1 is essential in vesicle trafficking from the Golgi apparatus to the endoplasmic reticulum (ER) through an interaction with Zeste-White 10 (ZW10). We have isolated a novel interaction between Rint1 and the human papillomavirus 16 (HPV16) transcription and replication factor E2. E2 binds to Rint1 within its ZW10 interaction domain, and we show that in the absence of E2, Rint1 is localized to the ER and associates with ZW10. E2 expression results in a disruption of the Rint1-ZW10 interaction and an accumulation of nuclear Rint1, coincident with a significant reduction in vesicle movement from the ER to the Golgi apparatus. Interestingly, nuclear Rint1 and members of the Mre11/Rad50/Nbs1 (MRN) complex were found in distinct E2 nuclear foci, which peaked during mid-S phase, indicating that the recruitment of Rint1 to E2 foci within the nucleus may also result in the recruitment of this DNA damage-sensing protein complex. We show that exogenous Rint1 expression enhances E2-dependent virus replication. Conversely, the overexpression of a truncated Rint1 protein that retains the E2 binding domain but not the Rad50 binding domain acts as a dominant negative inhibitor of E2-dependent HPV replication. Put together, these experiments demonstrate that the interaction between Rint1 and E2 has an important function in HPV replication.IMPORTANCEHPV infections are an important driver of many epithelial cancers, including those within the anogenital and oropharyngeal tracts. The HPV life cycle is tightly regulated and intimately linked to the differentiation of the epithelial cells that it infects. HPV replication factories formed in the nucleus are locations where viral DNA is copied to support virus persistence and amplification of infection. The recruitment of specific cellular protein complexes to these factories aids efficient and controlled viral replication. We have identified a novel HPV-host interaction that functions in the cellular response to DNA damage and cell cycle control. We show that the HPV E2 protein targets Rad50-interacting protein 1 (Rint1) to facilitate virus genome replication. These findings add to our understanding of how HPV replicates and the host cell pathways that are targeted by HPV to support virus replication. Understanding these pathways will allow further research into novel inhibitors of HPV genome replication.
authors.affiliation: University of Birmingham, Institute of Cancer and Genomic Sciences, Birmingham, United Kingdom.
authors.forename: Karen
authors.initials: K
authors.lastname: Campos-León
authors.affiliation: University of Birmingham, Institute of Cancer and Genomic Sciences, Birmingham, United Kingdom.
authors.forename: Kalpanee
authors.initials: K
authors.lastname: Wijendra
authors.affiliation: University of Birmingham, Institute of Cancer and Genomic Sciences, Birmingham, United Kingdom.
authors.forename: Abida
authors.initials: A
authors.lastname: Siddiqa
authors.affiliation: University of Birmingham, Institute of Cancer and Genomic Sciences, Birmingham, United Kingdom.
authors.forename: Ieisha
authors.initials: I
authors.lastname: Pentland
authors.affiliation: University of Birmingham, Institute of Cancer and Genomic Sciences, Birmingham, United Kingdom.
authors.forename: Katherine M
authors.initials: KM
authors.lastname: Feeney
authors.affiliation: University of St Andrews, School of Medicine, St Andrews, United Kingdom.
authors.forename: Alison
authors.initials: A
authors.lastname: Knapman
authors.affiliation: University of St Andrews, School of Medicine, St Andrews, United Kingdom.
authors.forename: Rachel
authors.initials: R
authors.lastname: Davies
authors.affiliation: Indiana University School of Medicine, Department of Dermatology, Indianapolis, Indiana, USA.
authors.forename: Elliot J
authors.identifier: http://orcid.org/0000-0002-8104-0703
authors.initials: EJ
authors.lastname: Androphy
authors.affiliation: University of Birmingham, Institute of Cancer and Genomic Sciences, Birmingham, United Kingdom j.l.parish@bham.ac.uk.
authors.forename: Joanna L
authors.initials: JL
authors.lastname: Parish
mesh_terms: D000595:Amino Acid Sequence; D018797:Cell Cycle Proteins / Q000378:metabolism*; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis; D004268:DNA-Binding Proteins / Q000378:metabolism*; D052162:Human papillomavirus 16 / Q000502:physiology*; D006801:Humans; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009856:Oncogene Proteins, Viral / Q000378:metabolism*; D011485:Protein Binding; D000072756:Protein Conformation, alpha-Helical; D054730:Protein Interaction Domains and Motifs; D021381:Protein Transport; D018741:Replication Origin; D059807:S Phase Cell Cycle Checkpoints; D014779:Virus Replication
chemical_list: D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D004279:DNA, Viral; D004268:DNA-Binding Proteins; C054855:E2 protein, Human papillomavirus type 16; D008869:Microtubule-Associated Proteins; D009856:Oncogene Proteins, Viral; C422909:RINT1 protein, human; C488261:ZW10 protein, human
keywords: E2; Rad50-interacting protein; papillomavirus; replication