Nucleic Acids Research, 1994, Vol. 22, No. 22 4583-4590
© 1994
MOLECULAR BIOLOGY |
DNA substrate specificity and cleavage kinetics of an archaeal homing-type endonuclease from Pyrobaculum organotrophum
Institute of Molecular Biology, Copenhagen Uniersity Sølvgade 83H, DK-1307 Copendhagen K, Denmark
*To whom correspondence should be addressed
Received August 25, 1994. Accepted October 10, 1994.
The protein encoded by intron 1 of the single 23S rRNA gene of the archaeal hyperthermophile Pyrobaculum organotrophum was isolated and shown to constitute a homing-type DNA endonuclease, l-Porl. It cleaves the intron- 23S rDNA of the closely related organism Pyrobaculum islandicum near the site of intron insertion in Pb.organotrophum. In contrast, no endonuclease activity was detected for the protein product of intron 2 of the same gene of Pb.organotrophum which, like l-Porl, carries the LAGLI-DADG motif, common to group I intron-encoded homing enzymes. I-Porl cleaves optimally at 80°C, with kcat and Kmvalues of about 2 min-1 and 4 nM, respectively, and generates four nucleotide 3'-overhangs and 5'-phosphates. It can cleave a 25 base pair DNA fragment encompassing the intron insertion site. A mutationselection study established the base pair specificity of the endonuclease within a 17 bp region, from positions –6 to +11 with respect to the intron-insertion site. Four of the essential base pairs encode the sequence involved in the intron-exon interaction in the pre-rRNA that is required for recognition by the RNA splicing enzymes. Properties of the enzyme are compared andcontrasted with those of eucaryotic homing endonucleases.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  J. Prieto, J.-C. Epinat, P. Redondo, E. Ramos, D. Padro, F. Cedrone, G. Montoya, F. Paques, and F. J. Blanco Generation and Analysis of Mesophilic Variants of the Thermostable Archaeal I-DmoI Homing Endonuclease J. Biol. Chem., February 15, 2008; 283(7): 4364 - 4374. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Nomura, Y. Morinaga, N. Shirai, and Y. Sako I-ApeI: a novel intron-encoded LAGLIDADG homing endonuclease from the archaeon, Aeropyrum pernix K1 Nucleic Acids Res., July 26, 2005; 33(13): e116 - e116. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Drouin, P. Lucas, C. Otis, C. Lemieux, and M. Turmel Biochemical characterization of I-CmoeI reveals that this H-N-H homing endonuclease shares functional similarities with H-N-H colicins Nucleic Acids Res., November 15, 2000; 28(22): 4566 - 4572. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Yu and M. R. Lieber The Nicking Step in V(D)J Recombination Is Independent of Synapsis: Implications for the Immune Repertoire Mol. Cell. Biol., November 1, 2000; 20(21): 7914 - 7921. [Abstract] [Full Text]
|
|
|
|
|
|
J. Lin and V. M. Vogt Functional {alpha}-fragment of {beta}-galactosidase can be expressed from the mobile group I intron PpLSU3 embedded in yeast pre-ribosomal RNA derived from the chromosomal rDNA locus Nucleic Acids Res., March 15, 2000; 28(6): 1428 - 1438. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Monteilhet, D. Dziadkowiec, T. Szczepanek, and J. Lazowska Purification and characterization of the DNA cleavage and recognition site of I-ScaI mitochondrial group I intron encoded endonuclease produced in Escherichia coli Nucleic Acids Res., March 1, 2000; 28(5): 1245 - 1251. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Pingoud, H. Thole, F. Christ, W. Grindl, W. Wende, and A. Pingoud Photocross-linking of the Homing Endonuclease PI-SceI to Its Recognition Sequence J. Biol. Chem., April 9, 1999; 274(15): 10235 - 10243. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. W. Armbruster and C. J. Daniels Splicing of Intron-containing tRNATrp by the Archaeon Haloferax volcanii Occurs Independent of Mature tRNA Structure J. Biol. Chem., August 8, 1997; 272(32): 19758 - 19762. [Abstract] [Full Text] [PDF]
|
|