Peptides Inhibitory for the Transcriptional Regulatory Function of Human Papillomavirus E2

Takuma Fujii, David Austin, David Guo, Srinivasan Srimatkandada, Tao Wang, Kaneyuki Kubushiro, Nobuo Masumoto, Katsumi Tsukazaki, Shiro Nozawa, Albert B. Deisseroth

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Purpose: Human papillomavirus (HPV) infections are associated with cervical neoplasia. Cellular and viral proteins are known to interact with the papillomavirus E2 protein to initiate transcription and DNA replication in the HPV life cycle. Our aim was to identify peptides that bind to the HPV16 E2 protein and thereby inhibit its ability to alter the transcriptional activity of other genes. Experimental Design: The HPV16 E2 protein was expressed and purified to near homogeneity in bacteria. We screened a phage display library of random peptides for ones that bound to HPV16 E2 protein. Among the isolated phage clones, we found that tryptophan-rich peptide sequences appeared repetitively in successive cycles of phage library panning. Replacement of the tryptophan amino acids in these dodecapeptides reduced the degree to which these peptides bound to the E2 protein. These E2-binding peptides were tested for their ability to inhibit the transcriptional regulatory function of E2 in a test cell line, which contained an E2 gene and a luciferase reporter gene driven by an E2-dependent transcriptional promoter. Results: Delivery of four of the E2 binding peptides into the intracellular compartment of the test cell line resulted in suppression of the E2-dependent luciferase expression. Deletion of the tryptophan residues from these peptides reduced their E2 binding and their ability to suppress E2-dependent luciferase expression in the test cell line. Conclusions: These results suggest a strategy for the development of chemical inhibitors of E2-dependent transcription of viral genes in HPV-infected cells as an approach to the therapy of chronic HPV infections.

Original languageEnglish
Pages (from-to)5423-5428
Number of pages6
JournalClinical Cancer Research
Volume9
Issue number14
Publication statusPublished - 01-11-2003
Externally publishedYes

Fingerprint

Peptides
Luciferases
Tryptophan
Papillomavirus Infections
Proteins
Cell Line
Bacteriophages
Peptide Library
Viral Genes
Viral Proteins
Life Cycle Stages
DNA Replication
Reporter Genes
Genes
Research Design
Clone Cells
Bacteria
Amino Acids
Neoplasms
Therapeutics

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

Cite this

Fujii, T., Austin, D., Guo, D., Srimatkandada, S., Wang, T., Kubushiro, K., ... Deisseroth, A. B. (2003). Peptides Inhibitory for the Transcriptional Regulatory Function of Human Papillomavirus E2. Clinical Cancer Research, 9(14), 5423-5428.
Fujii, Takuma ; Austin, David ; Guo, David ; Srimatkandada, Srinivasan ; Wang, Tao ; Kubushiro, Kaneyuki ; Masumoto, Nobuo ; Tsukazaki, Katsumi ; Nozawa, Shiro ; Deisseroth, Albert B. / Peptides Inhibitory for the Transcriptional Regulatory Function of Human Papillomavirus E2. In: Clinical Cancer Research. 2003 ; Vol. 9, No. 14. pp. 5423-5428.
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abstract = "Purpose: Human papillomavirus (HPV) infections are associated with cervical neoplasia. Cellular and viral proteins are known to interact with the papillomavirus E2 protein to initiate transcription and DNA replication in the HPV life cycle. Our aim was to identify peptides that bind to the HPV16 E2 protein and thereby inhibit its ability to alter the transcriptional activity of other genes. Experimental Design: The HPV16 E2 protein was expressed and purified to near homogeneity in bacteria. We screened a phage display library of random peptides for ones that bound to HPV16 E2 protein. Among the isolated phage clones, we found that tryptophan-rich peptide sequences appeared repetitively in successive cycles of phage library panning. Replacement of the tryptophan amino acids in these dodecapeptides reduced the degree to which these peptides bound to the E2 protein. These E2-binding peptides were tested for their ability to inhibit the transcriptional regulatory function of E2 in a test cell line, which contained an E2 gene and a luciferase reporter gene driven by an E2-dependent transcriptional promoter. Results: Delivery of four of the E2 binding peptides into the intracellular compartment of the test cell line resulted in suppression of the E2-dependent luciferase expression. Deletion of the tryptophan residues from these peptides reduced their E2 binding and their ability to suppress E2-dependent luciferase expression in the test cell line. Conclusions: These results suggest a strategy for the development of chemical inhibitors of E2-dependent transcription of viral genes in HPV-infected cells as an approach to the therapy of chronic HPV infections.",
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Fujii, T, Austin, D, Guo, D, Srimatkandada, S, Wang, T, Kubushiro, K, Masumoto, N, Tsukazaki, K, Nozawa, S & Deisseroth, AB 2003, 'Peptides Inhibitory for the Transcriptional Regulatory Function of Human Papillomavirus E2', Clinical Cancer Research, vol. 9, no. 14, pp. 5423-5428.

Peptides Inhibitory for the Transcriptional Regulatory Function of Human Papillomavirus E2. / Fujii, Takuma; Austin, David; Guo, David; Srimatkandada, Srinivasan; Wang, Tao; Kubushiro, Kaneyuki; Masumoto, Nobuo; Tsukazaki, Katsumi; Nozawa, Shiro; Deisseroth, Albert B.

In: Clinical Cancer Research, Vol. 9, No. 14, 01.11.2003, p. 5423-5428.

Research output: Contribution to journalArticle

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T1 - Peptides Inhibitory for the Transcriptional Regulatory Function of Human Papillomavirus E2

AU - Fujii, Takuma

AU - Austin, David

AU - Guo, David

AU - Srimatkandada, Srinivasan

AU - Wang, Tao

AU - Kubushiro, Kaneyuki

AU - Masumoto, Nobuo

AU - Tsukazaki, Katsumi

AU - Nozawa, Shiro

AU - Deisseroth, Albert B.

PY - 2003/11/1

Y1 - 2003/11/1

N2 - Purpose: Human papillomavirus (HPV) infections are associated with cervical neoplasia. Cellular and viral proteins are known to interact with the papillomavirus E2 protein to initiate transcription and DNA replication in the HPV life cycle. Our aim was to identify peptides that bind to the HPV16 E2 protein and thereby inhibit its ability to alter the transcriptional activity of other genes. Experimental Design: The HPV16 E2 protein was expressed and purified to near homogeneity in bacteria. We screened a phage display library of random peptides for ones that bound to HPV16 E2 protein. Among the isolated phage clones, we found that tryptophan-rich peptide sequences appeared repetitively in successive cycles of phage library panning. Replacement of the tryptophan amino acids in these dodecapeptides reduced the degree to which these peptides bound to the E2 protein. These E2-binding peptides were tested for their ability to inhibit the transcriptional regulatory function of E2 in a test cell line, which contained an E2 gene and a luciferase reporter gene driven by an E2-dependent transcriptional promoter. Results: Delivery of four of the E2 binding peptides into the intracellular compartment of the test cell line resulted in suppression of the E2-dependent luciferase expression. Deletion of the tryptophan residues from these peptides reduced their E2 binding and their ability to suppress E2-dependent luciferase expression in the test cell line. Conclusions: These results suggest a strategy for the development of chemical inhibitors of E2-dependent transcription of viral genes in HPV-infected cells as an approach to the therapy of chronic HPV infections.

AB - Purpose: Human papillomavirus (HPV) infections are associated with cervical neoplasia. Cellular and viral proteins are known to interact with the papillomavirus E2 protein to initiate transcription and DNA replication in the HPV life cycle. Our aim was to identify peptides that bind to the HPV16 E2 protein and thereby inhibit its ability to alter the transcriptional activity of other genes. Experimental Design: The HPV16 E2 protein was expressed and purified to near homogeneity in bacteria. We screened a phage display library of random peptides for ones that bound to HPV16 E2 protein. Among the isolated phage clones, we found that tryptophan-rich peptide sequences appeared repetitively in successive cycles of phage library panning. Replacement of the tryptophan amino acids in these dodecapeptides reduced the degree to which these peptides bound to the E2 protein. These E2-binding peptides were tested for their ability to inhibit the transcriptional regulatory function of E2 in a test cell line, which contained an E2 gene and a luciferase reporter gene driven by an E2-dependent transcriptional promoter. Results: Delivery of four of the E2 binding peptides into the intracellular compartment of the test cell line resulted in suppression of the E2-dependent luciferase expression. Deletion of the tryptophan residues from these peptides reduced their E2 binding and their ability to suppress E2-dependent luciferase expression in the test cell line. Conclusions: These results suggest a strategy for the development of chemical inhibitors of E2-dependent transcription of viral genes in HPV-infected cells as an approach to the therapy of chronic HPV infections.

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Fujii T, Austin D, Guo D, Srimatkandada S, Wang T, Kubushiro K et al. Peptides Inhibitory for the Transcriptional Regulatory Function of Human Papillomavirus E2. Clinical Cancer Research. 2003 Nov 1;9(14):5423-5428.