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PMID: 8335007 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Asymmetrical recognition and activity of the I-SceI endonuclease on its site and on intron-exon junctions.

The EMBO journal ·Vol. 12 ·No. 7 ·1993-07-00 ·Pages 2939-47

Perrin A, Buckle M, Dujon B

Abstract

Group I intron-encoded endonucleases represent a new class of double strand cutting endonucleases whose function is to initiate the homing of introns by generating double strand breaks in site-specific sequences. We have studied the mechanism of interaction of the I-SceI endonuclease with different DNA substrates derived from its natural site in the intron-less gene or from intron-exon junctions in the gene with an intron. We show that the enzyme recognizes its asymmetrical site with high affinity binding to the sequence corresponding to the downstream exon followed by binding to the upstream exon and catalysis of phosphodiester bond hydrolysis. Asymmetrical nicking activity is observed as an intermediate of the cleavage reaction. In the intron-containing gene, the enzyme recognizes the downstream intron-exon junction without any cleavage activity. This binding raises the possibility of a specific function of homing endonucleases in either gene expression or intron homing steps subsequent to DNA cleavage.

MeSH Terms
Base Sequence Cloning, Molecular DNA/metabolism Deoxyribonucleases, Type II Site-Specific/genetics,metabolism Escherichia coli Exons Introns Kinetics Molecular Sequence Data Saccharomyces cerevisiae Proteins Substrate Specificity
Chemicals
Saccharomyces cerevisiae Proteins DNA SCEI protein, S cerevisiae Deoxyribonucleases, Type II Site-Specific
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Perrin A
Département de Biologie Moléculaire, Institut Pasteur, Paris, France.
Buckle M
Dujon B
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1993-07-00
Pages
2939-47
Language
English
Region
England
NLM ID
8208664
PMCID
PMC413549
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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