PhD Scientific Days 2020

Molecular Sciences III. Lectures

Investigation of LCR Activity and E2 Viral Protein of Human Papillomavirus 11 Intratypic Variants

Előadó neve

Nagy, Zsófia

Előadó munkahelye

University of Debrecen, Faculty of Medicine, Department of Medical Microbiology

Előadó telefonszáma

+36308769859

Előadó e-mail címe

n.zsofia91@gmail.com

Az előadás címe

Investigation of LCR Activity and E2 Viral Protein of Human Papillomavirus 11 Intratypic Variants

Szerző(k) neve és munkahelye

Zsófia Nagy1, Attila Szűcs2, Krisztina Szarka1
1 Department of Medical Microbiology; 2 Clinic of Otorhinolaryngology and Head & Neck Surgery; Faculty of Medicine, University of Debrecen, Nagyerdei krt 98. Debrecen, Hungary

Szekció

Molecular Sciences III. Lectures

Language of the presentation

Hungarian

Section, first choice

Molecular Sciences

Section, second choice

Health Sciences

Összefoglaló szövege

Our former results showed that different severity of recurrent respiratory papillomatosis may be associated with given nucleotide polymorphisms identified in the ORF encoding replication and transcription regulatory protein E2 and in the long control region (LCR) of human papillomavirus virus (HPV) 11.
Our aim was to determine the interaction of LCR and E2 viral protein by transient co-transfection experiments to analyse how the polymorphisms in LCR and E2 influence LCR activity.
Six E2 variants (V1-V6) and six LCR patterns (P1-P6) were identified by comparison with HPV11 reference sequence (M14119). The P1 LCR pattern and the V1 E2 variant were present in nine HPV11 sequences of our samples. The other five HPV11 sequence carried unique LCR pattern and E2 variant. The identified unique LCRs were cloned into pALuc reporter vector and the E2 ORFs were cloned into pCDNA3.1+ expression vector, then transformed into Escherichia coli XL-1 bacterial strain. Plasmids transfected into HEP-2 cells with Lipofectamine 2000. Luciferase activities were compared by ANOVA with Tukey’s post-test.
In variant E2 and reference LCR co-transfection experiments, the highest luciferase activity was measured for reference LCR and E2 combination and in case of an identical HPV11 genome originated from cervical atypia. In case of five E2 variants (V1; V2; V3; V4; V5), the LCR activity was increased significantly lower level than in case of the reference E2. For three unique combinations (V4 + P4, V1 + P2, V2 + P1) the transactivating effect of LCR was significantly lower than the reference. Analysing the effect of E2 variants on the own LCR patterns, highest activity was also observed for the combination of the reference LCR and the reference E2. In case of co-transfection of unique LCR patterns and reference E2, the highest activity was found to be the combination of P6 LCR+ reference E2. The activity of P1 LCR+ reference E2 and P2 LCR + reference E2 was significantly lower than all other combinations. The 58 bp deletion in V2 E2 variant significantly reduces the E2 activity in both reference LCR and unique LCR assays.
Our data suggest that individual HPV11 E2 variants influence the transactivation activity of HPV11 LCR sequences and seem to confirm our hypothesis, that intratypic variantion of HPV11 genome may influence the disease outcome.

Additional Information

Krisztina Szarka
szkrisz@med.unideb.hu

Bemutatás módja

Szóbeli

Kind

Szabad

Status

elfogadva

Accepted presentation method

szóbeli

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

4731

Start

10:40

End

10:55

Authors (legacy)

Zsófia Nagy1, Attila Szűcs2, Krisztina Szarka1
1 Department of Medical Microbiology; 2 Clinic of Otorhinolaryngology and Head & Neck Surgery; Faculty of Medicine, University of Debrecen, Nagyerdei krt 98. Debrecen, Hungary