Document Type

Article

Publication Date

2007

MeSH Terms

Inteins, Proteins, Endonucleases

Subject: LCSH

Proteins, Endonucleases

Disciplines

Biology | Ecology and Evolutionary Biology

Abstract

Homing endonucleases are site-specific and rare cutting endonucleases often encoded by intron or intein containing genes. They lead to the rapid spread of the genetic element that hosts them by a process termed ‘homing’; and ultimately the allele containing the element will be fixed in the population. PI-SceI, an endonuclease encoded as a protein insert or intein within the yeast V-ATPase catalytic subunit encoding gene (vma1), is among the best characterized homing endonucleases. The structures of the Sce VMA1 intein and of the intein bound to its target site are known. Extensive biochemical studies performed on the PI-SceI enzyme provide information useful to recognize critical amino acids involved in self-splicing and endonuclease functions of the protein. Here we describe an insertion of the Green Fluorescence Protein (GFP) into a loop which is located between the endonuclease and splicing domains of the Sce VMA1 intein. The GFP is functional and the additional GFP domain does not prevent intein excision and endonuclease activity. However, the endonuclease activity of the newly engineered protein was different from the wild-type protein in that it required the presence of Mn2+ and not Mg2+ metal cations for activity.

Comments

Unless stated otherwise, articles in International Journal of Biological Sciences are distributed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. Reproduction is permitted for personal, noncommercial use only, provided that the article is in whole, unmodified, and properly cited. For commercial or any other use, please request permission to reproduce the materials.

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

Publisher Citation

Senejani, A.G. and Gogarten, J.P. “Structural stability and endonuclease activity of PI-SceI GFP-fusion protein.” Int J Biol Sci 2007; 3:205-211

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