E2 protein is the major determinant of specificity at the human papillomavirus origin of replication
Autoři:
Airiin Laaneväli aff001; Mart Ustav aff001; Ene Ustav aff001; Marko Piirsoo aff001
Působiště autorů:
Institute of Technology, University of Tartu, Tartu, Tartumaa, Estonia
aff001; Icosagen Cell Factory Ltd., Õssu, Kambja, Tartumaa, Estonia
aff002; Estonian Academy of Sciences, Tallinn, Harjumaa, Estonia
aff003
Vyšlo v časopise:
PLoS ONE 14(10)
Kategorie:
Research Article
doi:
https://doi.org/10.1371/journal.pone.0224334
Souhrn
The replication of human papillomavirus (HPV) genomes requires E1 and E2 proteins as the viral trans-factors and the replication origin, located in the URR, as a cis-element. The minimal requirements for an HPV replication origin vary among different virus types but always include one or more binding sites for the E2 protein. The requirements for an E1 binding site seem to vary among different HPV genera, with alpha-HPV11 and -18 minimal origins able to replicate without E1 binding site in contrast to beta-HPV8. In the present article, we analysed the sequence requirements for the beta-HPV5 minimal origin of replication. We show that the HPV5 URR is able to replicate in U2OS cells without the sequence proposed as an E1 binding site, albeit at lower levels than wt URR, given that three E2 binding sites are intact and both viral replication proteins are present. The lack of an absolute requirement of the E1 binding site for the origin of replication of HPV5 led us to analyse whether the viral E1 and E2 proteins from other HPV types are competent to support replication from this origin. Surprisingly, the E1 and E2 proteins from beta-HPV types support replication from the origin in contrast to proteins from alpha-HPV types 11, -16, or -18. Furthermore, the replication proteins E1 and E2 of these alpha-HPV types are unable to support the replication of HPV5 URR, even if the E1 binding site is intact. In light of these results, we performed a detailed analysis of the ability of different combinations of E1 and E2 proteins from various alpha- and beta-HPV types to support the replication of URR sequences from the respective HPV types in the U2OS cell line.
Klíčová slova:
DNA replication – DNA-binding proteins – Human papillomavirus – Plasmid construction – Plasmid vectors – Transfection – Viral replication – HPV-11
Zdroje
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