Mapping of Betapapillomavirus Human Papillomavirus 5 Transcription and Characterization of Viral-Genome Replication Function

Sankovski, Eve; Männik, Andres; Geimanen, Jelizaveta; Ustav, Ene; Ustav, Mart

doi:10.1128/jvi.01841-13

Cited by 44 publications

(67 citation statements)

References 61 publications

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“…Analysis of the MnPV TSSs revealed that the viral early promoter is located upstream of the E6 gene, at nt 78. This early promoter contains a TATA box 23 bp upstream of the TSS, in accordance with other early TSSs found upstream of E6 in other PVs like HPV18 P102 (Wang et al, 2011), HPV16 P97 (Grassmann et al, 1996), HPV5 P191 (Sankovski et al, 2014), HPV8 P175 (Stubenrauch et al, 1992) and HPV31 P99 (Hummel et al, 1992). This early TSS is very homogeneous, with all transcripts beginning at exactly the same nucleotide.…”

Section: Discussionsupporting

confidence: 81%

“…This is in contrast to data from other extensively studied PVs, like HPV18 (Wang et al, 2011) and HPV16 (Grassmann et al, 1996), for which a heterogeneous early TSS was found, differing in a few nucleotides from the main TSS. Sankovski et al (2014), who recently published the HPV5 transcriptome, also found a homogeneous early TSS, SJs are identified by their splicing donor/acceptor position. The number of reads aligning to each junction as determined by Tophat2 is displayed in the column 'Reads'.…”

Section: Discussionmentioning

confidence: 93%

“…This element does not conform to the canonical TATA box but is located in a favoured position; experimental confirmation that this element is functional is needed. A late differentiation-dependent promoter, generally located within the E7 ORF, is also found in other PVs such as HPV5, HPV11, HPV16, HPV18 and HPV31 (Hummel et al, 1992;Stubenrauch et al, 1992;Grassmann et al, 1996;Wang et al, 2011;Sankovski et al, 2014;IsokPaas et al, 2015). Generally these are TATA-less promoters, although a putative TATA-box was found in the case of HPV5 P840, as in the case of our findings for MnPV.…”

Section: Discussionmentioning

confidence: 98%

“…We propose an alternative explanation for the differences found between the early TSSs from HPV5 and MnPV and those from the rest of the PVs studied, based on the method used for their detection. Both Sankovski et al (2014) and our study used the 'First Choice RLM-RACE Kit', which is designed to amplify cDNA only from capped mRNA, usually producing a homogeneous product after PCR. As a matter of fact, Similar transcripts were found in several PVs in the published database for papillomavirus transcript maps (Baker & Calef, 1997), and also in particular in HPV5 (Sankovski et al, 2014), HPV11 (Chiang et al, 1991), HPV 16 (Sherman & Alloul, 1992;Milligan et al, 2007;Chen et al, 2014) and HPV18 (Wang et al, 2011).…”

Section: Discussionmentioning

confidence: 99%

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Transcriptome analysis of Mastomys natalensis papillomavirus in productive lesions after natural infection

Salvermoser

Chotewutmontri

Braspenning-Wesch

et al. 2016

Journal of General Virology

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Mastomys coucha, an African rodent, is a useful animal model of papillomavirus infection, as it develops both premalignant and malignant skin tumors as a consequence of a persistent infection with Mastomys natalensis papillomavirus (MnPV). In this study, we mapped the MnPV transcriptome in productive lesions by both classical molecular techniques and high-throughput RNA sequencing. Combination of these methods revealed a complex and comprehensive transcription map, with novel splicing events not described in other papillomaviruses. Furthermore, these splicing occurrences could potentially lead to the expression of novel E2, E1^E4, E7 and L2 isoforms. Expression level estimation of each transcript showed that lateregion mRNAs considerably outnumber early transcripts, with species coding for L1 and E1^E4 being the most abundant. In summary, the full transcription map assembled in this study will allow us to further understand MnPV gene expression and the mechanisms that lead to natural tumour development.

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Section: Discussionsupporting

confidence: 81%

Section: Discussionmentioning

confidence: 93%

Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

See 2 more Smart Citations

Transcriptome analysis of Mastomys natalensis papillomavirus in productive lesions after natural infection

Salvermoser

Chotewutmontri

Braspenning-Wesch

et al. 2016

Journal of General Virology

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“…In these differentiated IFI16-silenced NIKS-HPV18 cells, viral-load values were significantly increased compared with differentiated control cells. The second model consisted of U2OS cells transfected by electroporation with HPV18 minicircles, which sustain high levels of viral replication without the need for any differentiation stimuli (40,57), thus providing a suitable model for the assessment of IFI16 antiviral activity. In both cell lines, viral genomes were established as stably replicating extrachromosomal plasmids, as demonstrated by Southern blotting.…”

Section: Discussionmentioning

confidence: 99%

The Nuclear DNA Sensor IFI16 Acts as a Restriction Factor for Human Papillomavirus Replication through Epigenetic Modifications of the Viral Promoters

Cigno

Andrea

Borgogna

et al. 2015

J Virol

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The human interferon-inducible IFI16 protein, an innate immune sensor of intracellular DNA, was recently demonstrated to act as a restriction factor for human cytomegalovirus (HCMV) and herpes simplex virus 1 (HSV-1) infection by inhibiting both viral-DNA replication and transcription. Through the use of two distinct cellular models, this study provides strong evidence in support of the notion that IFI16 can also restrict human papillomavirus 18 (HPV18) replication. In the first model, an immortalized keratinocyte cell line (NIKS) was used, in which the IFI16 protein was knocked down through the use of small interfering RNA (siRNA) technology and overexpressed following transduction with the adenovirus IFI16 (AdVIFI16) vector. The second model consisted of U2OS cells transfected by electroporation with HPV18 minicircles. In differentiated IFI16-silenced NIKS-HPV18 cells, viral-load values were significantly increased compared with differentiated control cells. Consistent with this, IFI16 overexpression severely impaired HPV18 replication in both NIKS and U2OS cells, thus confirming its antiviral restriction activity. In addition to the inhibition of viral replication, IFI16 was also able to reduce viral transcription, as demonstrated by viralgene expression analysis in U2OS cells carrying episomal HPV18 minicircles and HeLa cells. We also provide evidence that IFI16 promotes the addition of heterochromatin marks and the reduction of euchromatin marks on viral chromatin at both early and late promoters, thus reducing both viral replication and transcription. Altogether, these results argue that IFI16 restricts chromatinized HPV DNA through epigenetic modifications and plays a broad surveillance role against viral DNA in the nucleus that is not restricted to herpesviruses. IMPORTANCEIntrinsic immunity is mediated by cellular restriction factors that are constitutively expressed and active even before a pathogen enters the cell. The host nuclear factor IFI16 acts as a sensor of foreign DNA and an antiviral restriction factor, as recently demonstrated by our group for human cytomegalovirus (HCMV) and herpes simplex virus 1 (HSV-1). Here, we provide the first evidence that IFI16 inhibits HPV18 replication by repressing viral-gene expression and replication. This antiviral restriction activity was observed in immortalized keratinocytes transfected with the religated genomes and in U2OS cells transfected with HPV18 minicircles, suggesting that it is not cell type specific. We also show that IFI16 promotes the assembly of heterochromatin on HPV DNA. These changes in viral chromatin structure lead to the generation of a repressive state at both early and late HPV18 promoters, thus implicating the protein in the epigenetic regulation of HPV gene expression and replication. Many recent studies point to the importance of cell-type-and host-specific expression of antiviral factors in limiting viral infection (1-4). Some of the very early antiviral responses include the activation of intrinsic restriction factors at h...

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Construction of a Transcription Map for Papillomaviruses using RACE, RNase Protection, and Primer Extension Assays

Wang

Zheng

2016

CP Microbiology

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Papillomaviruses are a family of small, non-enveloped DNA tumor viruses. Knowing a complete transcription map of each papillomavirus genome can provide guidance for various papillomavirus studies. This unit provides detailed protocols to construct a transcription map of human papillomavirus type 18. The same approach can be easily adapted to other transcription map studies of any other papillomavirus genotype due to the high degree of conservation in genome structure, organization, and gene expression among papillomaviruses. The focused methods are 5-and 3-rapid amplification of cDNA ends (RACE), which are techniques commonly used in molecular biology to obtain fulllength RNA transcript or to map a transcription start site (TSS) or an RNA polyadenylation (pA) cleavage site. Primer walking RT-PCR is a method for studying the splicing junction of RACE products. In addition, RNase protection assay and primer extension are also introduced as alternative methods in the mapping analysis.

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Mapping of Betapapillomavirus Human Papillomavirus 5 Transcription and Characterization of Viral-Genome Replication Function

Cited by 44 publications

References 61 publications

Transcriptome analysis of Mastomys natalensis papillomavirus in productive lesions after natural infection

Transcriptome analysis of Mastomys natalensis papillomavirus in productive lesions after natural infection

The Nuclear DNA Sensor IFI16 Acts as a Restriction Factor for Human Papillomavirus Replication through Epigenetic Modifications of the Viral Promoters

Construction of a Transcription Map for Papillomaviruses using RACE, RNase Protection, and Primer Extension Assays

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