Effect of E2 and long control region polymorphisms on disease severity in human papillomavirus type 11 mediated mucosal disease: Protein modelling and functional analysis

• Published in: Infection, genetics and evolution Vol. 93; p. 104948
• Main Authors: Nagy, Zsófia, Pethő, Zoltán, Kardos, Gábor, Major, Tamás, Szűcs, Attila, Szarka, Krisztina
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.09.2021
• Subjects: Condylomata Acuminata - virology; Female; Genome polymorphisms; Human papillomavirus 11 - genetics; Humans; Long control region; Low-risk human papillomavirus; Male; Papilloma - virology; Papillomavirus Infections - virology; Polymorphism, Genetic; Recurrent respiratory papillomatosis; Respiratory Tract Infections - virology; Severity of Illness Index; Transactivating potential; Viral Proteins - genetics; Transactivating potential; Low-risk human papillomavirus; Long control region; Recurrent respiratory papillomatosis; Genome polymorphisms
• ISSN: 1567-1348; 1567-7257
• DOI: 10.1016/j.meegid.2021.104948

Interaction of the long control region (LCR) and the E2 protein of HPV11s was studied by in silico modelling and in vitro functional analysis. Genomes of HPV11s from fifteen (six known and nine novel) patients (two solitary papillomas, eleven respiratory papillomatoses of different severity, one condyloma acuminatum and one cervical atypia) were sequenced; E2 polymorphisms were analysed in silico by protein modelling. E2 and LCR variants were cloned into pcDNA3.1+ expression vector and into pALuc reporter vector, respectively, transfected to HEp2 cells alone or in different combinations and the luciferase activity was measured. In the E2, the ubiquitous polymorphism K308R caused stronger binding between the dimers but did not alter DNA binding; E2s with this polymorphism were significantly less efficient than the reference in promoting LCR activity. The unique polymorphism Q86K changed the negative surface charge of E2 (Q86) to positive (K86). The unique polymorphisms S245F and N247T in the hinge region disrupt a probable phosphorylation site in a RXXS motif targeted by protein kinase A and B, but do not affect directly the amino acids critical to nuclear transport. Both unique patterns partly restored the LCR activating potential disrupted by K308R. A unique E2/E4 ORF with a 58-bp deletion leading to a frameshift and an early stop codon resulted in a practically nonfunctional E2, and was associated with a papillomatosis with dysplasia. When testing existing LCR-E2 combinations, LCR with intrinsically lower enhancer capacity was only marginally activated by its E2 (R308 and the deletion mutant), and did not significantly exceed the activity of the reference LCR without E2. Combined with more potent LCRs associated with more severe disease, the activity was significantly higher, but still significantly lower than LCRs with reference E2. In summary, LCR-E2 interaction determined by their polymorphisms may explain, at least partly, differences in disease severity. •Besides the reference, five LCR patterns and four E2 variants were identified.•HPV11 long control regions differ in their intrinsic transactivating potential.•E2s with the ubiquitous K308R change dimerize easier but enhance LCR less strongly.•The prototype HPV11 sequence represents a unique variant with high virulence.•Interaction of E2 and LCR determines transactivation and contributes to virulence.