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Journals / The Journal of Turkish Phytopathology / 2012 / Cilt: 41 - Sayı: 1-2-3

Cloning and Sequencing of Virus Inhibiting Gene Encoding an Antiviral Protein From the Leaves of Pokeweed (Phytolacca Americana L.)

Hikmet Murat SİPAHİOĞLU
İlhan KAYA
Mustafa USTA
Sibel SAMSUN KIZILDAĞ
Pages
19–26
Publication Date
2014-05-28
DOI
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AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

Sequence analysis of PAP-I revealed that it contains a single point mutation, changing the Leucine (L) at position 273 to Phenylalanine (F) (L273F) at the putative active site.

📌 3 Key Takeaways

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Objective & Hypothesis:Pokeweed antiviral protein (PAP) is single-chain ribosome-inactivating protein (RIP) of Phytolacca americana L. (Pokeweed) that is characterized by its ability to depurinate ribosomes.
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Key Findings & Data:In the present study, we cloned and sequenced complete gene of pokeweed antiviral protein type 1 (PAP-I) from the summer leaves of pokeweed collected from Trabzon (Turkey) using a pair of gene specific primers based on the known N- and C-terminal nucleotide sequences of PAP gene.
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Conclusion & Impact:Sequence analysis of PAP-I revealed that it contains a single point mutation, changing the Leucine (L) at position 273 to Phenylalanine (F) (L273F) at the putative active site.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:antiviral proteins · characterization · cloning

Abstract

Pokeweed antiviral protein (PAP) is single-chain ribosome-inactivating protein (RIP) of Phytolacca americana L. (Pokeweed) that is characterized by its ability to depurinate ribosomes. In the present study, we cloned and sequenced complete gene of pokeweed antiviral protein type 1 (PAP-I) from the summer leaves of pokeweed collected from Trabzon (Turkey) using a pair of gene specific primers based on the known N- and C-terminal nucleotide sequences of PAP gene. A product of 942 base pair was purified and inserted into pGEM-T Easy vector (Promega), downstream of the T7 promoter, and transformed into E. coli strain, JM 109. The PAP-I cistron of P. americana contained 313 amino acid residues. The DNA sequence of 942 base pairs included an open reading frame (ORF). The nucleotide sequences of PAP-I gene contained no introns and the comparison with the PAP-I sequence with the PAP isoform, showed an identity of 80-99%. Sequence analysis of PAP-I revealed that it contains a single point mutation, changing the Leucine (L) at position 273 to Phenylalanine (F) (L273F) at the putative active site.

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Cite This Article

SİPAHİOĞLU, H. M., KAYA, İ., USTA, M., KIZILDAĞ, S. S. (2014). Cloning and Sequencing of Virus Inhibiting Gene Encoding an Antiviral Protein From the Leaves of Pokeweed (Phytolacca Americana L.). The Journal of Turkish Phytopathology, Cilt: 41 - Sayı: 1-2-3, 19–26. https://ajindex.com/articles/844293

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References (10)

  1. Aron, G.M. and Irvin, J.D. 1980. Inhibition of herpes simplex virus multiplication by the pokeweed antiviral protein. Antimicrobial Agents and Chemotherapy.17:1032–1033.
  2. Barbieri, L.,Valbonesi, P., Bonora, E., Gorini, P., Bolognesi, A. and Stirpe, F. 1997. Polynucleotide:adenosine glycosidase activity of ribosome-inactivating proteins: effect on DNA, RNA and poly (A). Nucleic Acids Research. 25:518–522.
  3. Bijal, A., Parikh, C.C. and Tumer, N.E. 2002. Pokeweed Antiviral protein regulates the stability of its own mRNA by a mechanism that requires depurination but can be separated from depurination of the alpha-sarcin/ricin loop of rRNA. The Journal of Biological Chemistry. 277:41428–41437.
  4. Dore, J. M., Gras, E., Depierre, F. and Wijdens, J. 1993. Mutations dissociating the inhibitory activity of the pokeweed antiviral protein on eukaryote translation and Escherichia coli growth. Nucleic Acids Res.21, 4200–4205.
  5. Endo, Y., Tsurugi, K.and Lambert, J.M. 1988. The site of action of six different ribosome-inactivating proteins from plants on eukaryotic ribosomes: the RNA N-glycosidase activity of the proteins. Biochemical and Biophysical Research Communications.150:1032–1036. Foà-Tomasi, L., Campadelli-Fiume, G., Barbieri, L.and Stirpe, F. 1982. Effect of ribosome-inactivating proteins on virus-infected cells. Inhibition of virus multiplication and of protein synthesis. Archives of Virology. 71 (4):323–332.
  6. Hartley, M.R., Legname, G., Osborn, R., Chen, Z.and Lord, J.M. 1991.Single-chain ribosome inactivating proteins from plants depurinate Escherichia coli 23S ribosomal RNA. FEBS Letters.290: 65–68.
  7. Houston, L.L., Ramakrishnanz, S.and Hermodsong, M.A. 1983. Seasonal variations in different forms of pokeweed antiviral protein a potent inactivator of ribosomes. Journal of Biological Chemistry. 258:9601–9604.
  8. Hudak, K.A., Dinman, J.D. andTumer, N.E. 1999. Pokeweed antiviral protein accesses ribosomes by binding to L3. Journal of Biological Chemistry. 274:3859–3864.
  9. Gendron, Y., and Kassanis, B. 1954. The importance of the host species in determining the action of virus inhibitors. Irvin, J.D.andUckun, F.M. 1992.Pokeweed antiviral protein: ribosome inactivation and therapeutic applications. Pharmacology & Therapeutics55:279–302.
  10. Kataoka, J., Habuka,N., Furruno, M., Miyano, M., Takanami,Y.and Koiwai, A. 1991. A sequence of Mirabilis antiviral protein (MAP), a ribosome inactivating protein with an antiviral property, from Mirablis jalapa L. andits expression in E. coli. J