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Plos One : I-pfop3I ; a Novel Nicking Hnh Homing Endonuclease Encoded in the Group I Intron of the Dna Polymerase Gene in Phormidium Foveolarum Phage Pf-wmp3, Volume 7

By Xu, Shuang-yong

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Book Id: WPLBN0003939857
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Reproduction Date: 2015

Title: Plos One : I-pfop3I ; a Novel Nicking Hnh Homing Endonuclease Encoded in the Group I Intron of the Dna Polymerase Gene in Phormidium Foveolarum Phage Pf-wmp3, Volume 7  
Author: Xu, Shuang-yong
Volume: Volume 7
Language: English
Subject: Journals, Science, Medical Science
Collections: Periodicals: Journal and Magazine Collection
Historic
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Publisher: Plos

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Xu, S. (n.d.). Plos One : I-pfop3I ; a Novel Nicking Hnh Homing Endonuclease Encoded in the Group I Intron of the Dna Polymerase Gene in Phormidium Foveolarum Phage Pf-wmp3, Volume 7. Retrieved from http://netlibrary.net/


Description
Description : Homing endonucleases encoded in a group I self-splicing intron in a protein-coding gene in cyanophage genomes have not been reported, apart from some free-standing homing edonucleases. In this study, a nicking DNA endonuclease, I-PfoP3I, encoded in a group IA2 intron in the DNA polymerase gene of a T7-like cyanophage Pf-WMP3, which infects the freshwater cyanobacterium Phormidium foveolarum is described. The Pf-WMP3 intron splices efficiently in vivo and self-splices in vitro simultaneously during transcription. I-PfoP3I belongs to the HNH family with an unconventional C-terminal HNH motif. IPfoP3I nicks the intron-minus Pf-WMP3 DNA polymerase gene more efficiently than the Pf-WMP4 DNA polymerase gene that lacks any intervening sequence in vitro, indicating the variable capacity of I-PfoP3I. I-PfoP3I cleaves 4 nt upstream of the intron insertion site on the coding strand of EXON 1 on both intron-minus Pf-WMP3 and Pf-WMP4 DNA polymerase genes. Using an in vitro cleavage assay and scanning deletion mutants of the intronless target site, the minimal recognition site was determined to be a 14 bp region downstream of the cut site. I-PfoP3I requires Mg2+, Ca2+ or Mn2+ for nicking activity. Phylogenetic analysis suggests that the intron and homing endonuclease gene elements might be inserted in Pf- WMP3 genome individually after differentiation from Pf-WMP4. To our knowledge, this is the first report of the presence of a group I self-splicing intron encoding a functional homing endonuclease in a protein-coding gene in a cyanophage genome.

 

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