Identification of a new promoter in the early region of the human papillomavirus type 16 genome

Braunstein, Thomas Hartig; Madsen, B.S.; Gavnholt, Britta; Rosenstierne, Maiken Worsøe; Johnsen, Christina Koefoed; Norrild, Bodil

doi:10.1099/0022-1317-80-12-3241

Cited by 14 publications

(12 citation statements)

References 39 publications

(59 reference statements)

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“…In addition to the new initiation sites, we mapped transcripts from P97 to initiate at nt 97 and 95. We also found a cluster of transcription start sites present at around nt 480, which have been described previously as the promoter P480 (Grassmann et al, 1996;Braunstein et al, 1999). To verify the presence of transcription start sites around nt 441, we performed RNase protection assays with four different probes.…”

Section: Discussionsupporting

confidence: 67%

“…The different fragments were inserted into the promoter-and enhancerless pGL3-Basic or pGL3-Enhancer vectors (Promega), the latter contains the simian virus 40 (SV40) enhancer. pGL3-Basic constructs were all modified to reduce background from the vector, as described by Braunstein et al (1999). Site-directed mutagenesis of the nt 272-448 fragment was performed using primers containing two point mutations (59-gcatggacgctagccttttcttcaggacacagtggcttttgacagttaatacacctacgtaacaaatca-39 and 59-gcatggacgctagccttttcttcaggacacagtggcttttgacagtgaagacacctaatt-39; mutations are shown in bold).…”

Section: Methodsmentioning

confidence: 99%

“…The pGL3-Basic vector was modified further to reduce background activity, as described by Braunstein et al (1999). In these experiments, the HPV-16 fragments contained only the transcription start sites around nt 441 (Fig.…”

Section: Promoter Activity Of Luciferase Reporter Constructsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…In HPV-16, a transcription start site was identified in the E6 region at nt 480 but this was not characterized further (Grassmann et al, 1996). We have previously identified and characterized a new promoter, P542, with a transcription start site at nt 542 in front of the E7 ORF (Braunstein et al, 1999;Glahder et al, 2003).Our previous work indicated the existence of promoter activity located upstream of P542. This led us to search for additional transcription start sites in the HPV-16 E6 ORF that could contribute to the expression of E7.…”

mentioning

confidence: 99%

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Identification and characterization of a cluster of transcription start sites located in the E6 ORF of human papillomavirus type 16

Rosenstierne

Vinther

Hansen

et al. 2003

Journal of General Virology

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Human papillomavirus type 16 (HPV-16) is the prototype strain among the malignant types of HPV in the western world. The main promoter, P97, located in front of the E6 ORF, has been shown to control expression of the oncogenes E6 and E7. These oncogenes are expressed continuously in HPV-16-transformed cells. In contrast to malignant HPV types, non-malignant HPV types have separate promoters driving the expression of E6 and E7. Experiments have shown that the translation of E7 is more efficient from monocistronic than bicistronic transcripts encoding both E6 and E7. Here, identification of a cluster of transcription start sites located in the E6 ORF of HPV-16 is presented. Transcripts from this region contain the E7 ORF as the first reading frame. The cluster consists of multiple transcription start sites located around nt 441. Additional transcription start sites were identified in a cluster around nt 480. A transcription start site has been identified previously at nt 480 but has never been characterized further. The region responsible for transcription activity was mapped to nt 272-448. Mutational analysis showed that initiation of transcription is independent of a TATA-box element, which is consistent with the finding of multiple transcription start sites. Furthermore, it is shown that proteins from HeLa and SiHa nuclear cell extracts bind to the two regions at nt 291-314 and 388-411, and that these two regions influence transcription activity in a cell type-dependent manner. INTRODUCTIONHuman papillomaviruses (HPVs) are epitheliotropic DNA viruses with a double-stranded circular genome of approximately 8 kbp. During the last 20 years, more than 85 different types of HPV have been identified and characterized, and novel types are still being isolated (zur Hausen, 1999). HPV types are divided into malignant and nonmalignant types according to their ability to transform the host cell. The HPV genome consists of an early region encoding six to seven early proteins [E6, E7, E1, E2, E4 (E8) and E5], a late region encoding two late proteins (L1 and L2) and two regulatory regions, designated the long control region (LCR) and the small non-coding region (Seedorf et al., 1985). In the western world, HPV-16 is considered to be the prototype strain of the malignant types of HPV. The oncoproteins E6, E7 and E5 from malignant types of HPV are responsible for the transformation of the epithelial host cell. In the normal life cycle of the virus, oncoproteins induce unscheduled replication in differentiating keratinocytes to facilitate virus replication (for reviews, see zur Hausen, 1996; and Zwerschke & Jansen-Dürr, 2000). The E1 and E2 proteins are responsible for DNA replication (reviewed by Phelps et al., 1998) and E2 also functions as a transcriptional regulator (Nishimura et al., 2000; Tan et al., 1994;Ushikai et al., 1994). The E1^E4 protein binds to intermediate filaments and is believed to facilitate the release of virus particles. In addition, the E1^E4 protein is believed to be involved in post-transcription regu...

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Section: Discussionsupporting

confidence: 67%

Section: Methodsmentioning

confidence: 99%

Section: Promoter Activity Of Luciferase Reporter Constructsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

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Identification and characterization of a cluster of transcription start sites located in the E6 ORF of human papillomavirus type 16

Rosenstierne

Vinther

Hansen

et al. 2003

Journal of General Virology

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Identification of a novel promoter in the E2 open reading frame of the human papillomavirus type 18 genome

Kim

Taylor

2002

Journal of Medical Virology

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Northern blot and RT-PCR analyses indicated that the human papillomavirus E4 open reading frame is expressed in HeLa cells. Because integration at the E1 or E2 open reading frame would place the viral upstream regulatory region downstream of the viral late genes, the expression of E4 in HeLa cells is most likely regulated by host cellular promoter(s) or unidentified viral promoter(s) in the E2 region. Primer extension analysis and transient transfection experiments with luciferase reporter constructs under the transcriptional control of various subgenomic fragments of HPV-18 were carried out to identify and characterize functional promoters within the E2 region. The in vivo activity of a novel promoter located within the 5'-end region of the E2 open reading frame of human papillomavirus type 18 is demonstrated.

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New insights in Hippo signalling alteration in human papillomavirus-related cancers

Olmedo-Nieva

Muñoz-Bello

Manzo-Merino

et al. 2020

Cellular Signalling

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Identification of a new promoter in the early region of the human papillomavirus type 16 genome

Cited by 14 publications

References 39 publications

Identification and characterization of a cluster of transcription start sites located in the E6 ORF of human papillomavirus type 16

Identification and characterization of a cluster of transcription start sites located in the E6 ORF of human papillomavirus type 16

Identification of a novel promoter in the E2 open reading frame of the human papillomavirus type 18 genome

New insights in Hippo signalling alteration in human papillomavirus-related cancers

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