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PMID: 1475201 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The I-CeuI endonuclease recognizes a sequence of 19 base pairs and preferentially cleaves the coding strand of the Chlamydomonas moewusii chloroplast large subunit rRNA gene.

Nucleic acids research ·Vol. 20 ·No. 23 ·1992-12-11 ·Pages 6401-7

Marshall P, Lemieux C

Abstract

The I-CeuI endonuclease is a member of the growing family of homing endonucleases that catalyse mobility of group I introns by making a double-strand break at the homing site of these introns in cognate intronless alleles during genetic crosses. In a previous study, we have shown that a short DNA fragment of 26 bp, encompassing the homing site of the fifth intron in the Chlamydomonas eugametos chloroplast large subunit rRNA gene (Ce LSU.5), was sufficient for I-CeuI recognition and cleavage. Here, we report the recognition sequence of the I-CeuI endonuclease, as determined by random mutagenesis of nucleotide positions adjacent to the I-CeuI cleavage site. Single-base substitutions that completely abolish endonuclease activity delimit a 15-bp sequence whereas those that reduce the cleavage rate define a 19-bp sequence that extends from position -7 to position +12 with respect to the Ce LSU.5 intron insertion site. As the other homing endonucleases that have been studied so far, the I-CeuI endonuclease recognizes a non-symmetric degenerate sequence. The top strand of the recognition sequence is preferred for I-CeuI cleavage and the bottom strand most likely determines the rate of double-strand breaks.

MeSH Terms

Animals Base Composition Base Sequence Chlamydomonas/genetics,metabolism Chloroplasts/metabolism DNA Mutational Analysis DNA, Ribosomal/metabolism Endodeoxyribonucleases/metabolism Exons Kinetics Molecular Sequence Data Mutagenesis Mutagenesis, Site-Directed RNA, Ribosomal/genetics Substrate Specificity

Chemicals

DNA, Ribosomal RNA, Ribosomal Endodeoxyribonucleases endodeoxyribonuclease I-CeuI

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Marshall P

Département de Biochimie, Faculté des Sciences et de Génie, Université Laval, Québec, Canada.

Lemieux C

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33 references, click to expand

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Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/20.23.6401

Published

1992-12-11

Pages

6401-7

Language

English

Region

England

NLM ID

0411011

PMCID

PMC334533

Subset

External Links

PubMed source DOI: 10.1093/nar/20.23.6401 PMC Full Text View journal details

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The I-CeuI endonuclease recognizes a sequence of 19 base pairs and preferentially cleaves the coding strand of the Chlamydomonas moewusii chloroplast large subunit rRNA gene.

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