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Nucleic Acids Research, 2003, Vol. 31, No. 12 3071-3077
© 2003 Oxford University Press

The nicking homing endonuclease I-BasI is encoded by a group I intron in the DNA polymerase gene of the Bacillus thuringiensis phage Bastille

Markus Landthaler and David A. Shub

University at Albany, SUNY, Department of Biological Sciences and Center for Molecular Genetics, Albany, NY, USA

*To whom correspondence should be addressed. Tel: +1 518 442 4324; Fax: +1 518 442 4767; Email: shub{at}albany.edu
Present address:
Markus Landthaler, Rockefeller University, New York, NY, USA

Here we describe the discovery of a group I intron in the DNA polymerase gene of Bacillus thuringiensis phage Bastille. Although the intron insertion site is identical to that of the Bacillus subtilis phages SPO1 and SP82 introns, the Bastille intron differs from them substantially in primary and secondary structure. Like the SPO1 and SP82 introns, the Bastille intron encodes a nicking DNA endonuclease of the H-N-H family, I-BasI, with a cleavage site identical to that of the SPO1-encoded enzyme I-HmuI. Unlike I-HmuI, which nicks both intron-minus and intron-plus DNA, I-BasI cleaves only intron-minus alleles, which is a characteristic of typical homing endonucleases. Interestingly, the C-terminal portions of these H-N-H phage endonucleases contain a conserved sequence motif, the intron-encoded endonuclease repeat motif (IENR1) that also has been found in endonucleases of the GIY-YIG family, and which likely comprises a small DNA-binding module with a globular ßß{alpha}{alpha}ß fold, suggestive of module shuffling between different homing endonuclease families.


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