Characterization of the nuclear matrix targeting sequence (NMTS) of the BPV1 E8/E2 protein — the shortest known NMTS

Sankovski, Eve; Karro, Kristiina; Sepp, Mari; Kurg, Reet; Ustav, Mart; Abroi, Aare

doi:10.1080/19491034.2015.1074359

Cited by 5 publications

(6 citation statements)

References 60 publications

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“…The E1^M1 and E1^M2 in MmuPV1 might be similar to the E1Ma and E1M in HPV-11 [ 25 ]. The MmuPV1 E8^E2 gene product is likely similar to the E8^E2 gene products characterized in BPV-1 [ 41 – 44 ] and HPVs [ 45 , 46 ] and predicted to be encoded by most papillomaviruses [ 47 ]. Although the first AUG codon for each of the predicted coding regions has a strong Kozak sequence [ 20 ], we were unable to detect the expression of MmuPV1 E1, L2 and L1 proteins from a common eukaryotic expression vector in HEK293, MEK, 293FT or 293TT cells and unable to detect by Northern blotting a full-length RNA expressed from E1, L2 and L1 vector in HEK293 cells.…”

Section: Discussionmentioning

confidence: 96%

The full transcription map of mouse papillomavirus type 1 (MmuPV1) in mouse wart tissues

et al. 2017

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Mouse papillomavirus type 1 (MmuPV1) provides, for the first time, the opportunity to study infection and pathogenesis of papillomaviruses in the context of laboratory mice. In this report, we define the transcriptome of MmuPV1 genome present in papillomas arising in experimentally infected mice using a combination of RNA-seq, PacBio Iso-seq, 5’ RACE, 3’ RACE, primer-walking RT-PCR, RNase protection, Northern blot and in situ hybridization analyses. We demonstrate that the MmuPV1 genome is transcribed unidirectionally from five major promoters (P) or transcription start sites (TSS) and polyadenylates its transcripts at two major polyadenylation (pA) sites. We designate the P7503, P360 and P859 as “early” promoters because they give rise to transcripts mostly utilizing the polyadenylation signal at nt 3844 and therefore can only encode early genes, and P7107 and P533 as “late” promoters because they give rise to transcripts utilizing polyadenylation signals at either nt 3844 or nt 7047, the latter being able to encode late, capsid proteins. MmuPV1 genome contains five splice donor sites and three acceptor sites that produce thirty-six RNA isoforms deduced to express seven predicted early gene products (E6, E7, E1, E1^M1, E1^M2, E2 and E8^E2) and three predicted late gene products (E1^E4, L2 and L1). The majority of the viral early transcripts are spliced once from nt 757 to 3139, while viral late transcripts, which are predicted to encode L1, are spliced twice, first from nt 7243 to either nt 3139 (P7107) or nt 757 to 3139 (P533) and second from nt 3431 to nt 5372. Thirteen of these viral transcripts were detectable by Northern blot analysis, with the P533-derived late E1^E4 transcripts being the most abundant. The late transcripts could be detected in highly differentiated keratinocytes of MmuPV1-infected tissues as early as ten days after MmuPV1 inoculation and correlated with detection of L1 protein and viral DNA amplification. In mature warts, detection of L1 was also found in more poorly differentiated cells, as previously reported. Subclinical infections were also observed. The comprehensive transcription map of MmuPV1 generated in this study provides further evidence that MmuPV1 is similar to high-risk cutaneous beta human papillomaviruses. The knowledge revealed will facilitate the use of MmuPV1 as an animal virus model for understanding of human papillomavirus gene expression, pathogenesis and immunology.

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Section: Discussionmentioning

confidence: 96%

The full transcription map of mouse papillomavirus type 1 (MmuPV1) in mouse wart tissues

et al. 2017

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“…The highly conserved lysine at the second position (Fig. 5c) of the E8 motif is functionally important, and changing the lysine to an alanine eliminates the repressive function of E8^E2 in BPV1 (Sankovski et al, 2015). This E8 motif is likely the most ancestral and is approximately 150 million years old according to the divergence time of the PV genera encoding this type of E8 (Shah et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

“…In human papillomavirus 31 (HPV31), the E8^E2 protein represses transcription and extra-chromosomal replication, and many interactions responsible for these repressive functions have been described (Ammermann et al, 2008;Fertey et al, 2010;Powell et al, 2010). In addition to HPV31, the transcriptional repression activity of E8^E2 protein has been described for BPV1, Sylvilagus floridanus papillomavirus 1 (SfPV1), HPV16 and HPV5 (Choe et al, 1989;Jeckel et al, 2003;Lace et al, 2008Lace et al, , 2012Sankovski et al, 2014Sankovski et al, , 2015. In BPV1, the E8 peptide of the E8^E2 protein functions as a nuclear matrix-targeting signal (NMTS) (Sankovski et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

“…In addition to HPV31, the transcriptional repression activity of E8^E2 protein has been described for BPV1, Sylvilagus floridanus papillomavirus 1 (SfPV1), HPV16 and HPV5 (Choe et al, 1989;Jeckel et al, 2003;Lace et al, 2008Lace et al, , 2012Sankovski et al, 2014Sankovski et al, , 2015. In BPV1, the E8 peptide of the E8^E2 protein functions as a nuclear matrix-targeting signal (NMTS) (Sankovski et al, 2015). Repression of early gene expression appears to be a common property of E8^E2 in all PVs in which this protein has been described.…”

Section: Introductionmentioning

confidence: 99%

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Conservation of the E8 CDS of the E8^E2 protein among mammalian papillomaviruses

Puustusmaa

Abroi

2016

Journal of General Virology

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Papillomaviridae are small dsDNA viruses with a limited coding capacity. To fulfill all of the functional requirements for propagation and spreading, papillomaviruses use double coding and alternative protein isoforms. E8^E2 is an alternative E2 protein isoform that is generated by fusing the short E8 CDS that completely overlaps E1 to the 'hinge' and the DNA-binding region of the E2 protein via alternative transcription/splicing. The papillomaviruses in which E8^E2 mRNA sequences have been described exhibit a sparse phylogenomic distribution. Thus, it is not clear whether E8^E2 is an ancestral protein that has not been described for other papillomavirus types or whether it randomly appears because of the conservation of the E1 protein and occurs only coincidentally. We searched for potential E8 coding sequences in a non-redundant set of papillomaviruses and applied SynPlot2 and an in-house-developed algorithm (cRegions) to determine the most plausible of the above-mentioned scenarios. Beginning with nine experimentally described E8^E2 mRNAs, we predicted the potential E8 CDSs for more than 300 mammalian papillomavirus genomes. According to our analysis, E8^E2 is not a result of E1 coding and represents a protein in its own right, and it most likely has an ancestral origin that precedes the divergence of major mammalian papillomavirus genera.

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“…HEK 293TT cells were transfected with FLAG-HPV-31 E2 WT or Y102E and harvested 24 h later. Cell pellets were fractionated as previously described (47).…”

Section: Methodsmentioning

confidence: 99%

Human Papillomavirus 31 Tyrosine 102 Regulates Interaction with E2 Binding Partners and Episomal Maintenance

Gilson

Culleton

Xie

et al. 2020

J Virol

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Several serine and threonine residues of the papilloma virus early E2 protein have been found to be phosphorylated. By contrast, only one E2 tyrosine phosphorylation site in BPV-1 (tyrosine 102) and one HPV-16/31 (tyrosine 138) site have been characterized. Between BPV-1 and HPV-31 E2, 8 of the 11 tyrosines are conserved in the N-terminal domain, suggesting that phosphorylation of tyrosines has an essential role in E2 biology. In this paper we examine the effect of Y102 phosphorylation on HPV-31 E2 biology. Y102 proteins mutated either to the potential phospho-mimetic glutamic acid (Y102E) or to the non-phosphorylated homologue phenylalanine (Y102F) remain nuclear; however, Y102E is more associated with the nuclear matrix fraction. This is consistent with the inability of Y102E to bind TopBP1. Both BPV-1 and HPV-31 Y102E are similar in that neither bind the C-terminus of Brd4, but in all other aspects, the mutant behaves differently between the two families of papillomaviruses. BPV-1 Y102E was unable to bind E1 and did not replicate in a transient in-vitro assay, while HPV-31 Y102E binds E1 and replicated albeit at lower levels than wild type. To examine effect of E2 mutations under more native-like infection conditions, a neomycin selectable marker was inserted into L1/L2 of HPV-31 genome, creating HPV-31neo. This genome was maintained in every cell line tested for at least 50 days post-transfection/infection. Y102E in both transfection and infection conditions was unable to maintain high episome copy numbers in epithelial cell lines. IMPORTANCE Post-translational modifications by phosphorylation can change protein activities, binding partners, or localization. Tyrosine 102 is conserved between delta papillomavirus BPV-1 and alpha papillomavirus HPV-31 E2. We characterized mutations of HPV-31 E2 for interactions with relevant cellular binding partners and replication in the context of the viral genome.

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Characterization of the nuclear matrix targeting sequence (NMTS) of the BPV1 E8/E2 protein — the shortest known NMTS

Cited by 5 publications

References 60 publications

The full transcription map of mouse papillomavirus type 1 (MmuPV1) in mouse wart tissues

The full transcription map of mouse papillomavirus type 1 (MmuPV1) in mouse wart tissues

Conservation of the E8 CDS of the E8^E2 protein among mammalian papillomaviruses

Human Papillomavirus 31 Tyrosine 102 Regulates Interaction with E2 Binding Partners and Episomal Maintenance

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