Characterization of the cleavage site and the recognition sequence of the I-CreI DNA endonuclease encoded by the chloroplast ribosomal intron of Chlamydomonas reinhardtii.
The chloroplast ribosomal intron of Chlamydomonas reinhardtii encodes a sequence-specific DNA endonuclease (I-CreI), which is most probably involved in the mobility of this intron. Here we show that I-CreI generates a 4 bp staggered cleavage just downstream of the intron insertion site. The I-CreI recognition sequence is 19-24 bp in size and is located asymmetrically around the intron insertion site. Screening of natural variants of the I-CreI recognition sequence indicates that the I-CreI endonuclease tolerates single and even multiple base changes within its recognition sequence.
Dürrenberger
F
F
Department of Molecular Biology, University of Geneva, Switzerland.
Rochaix
J D
JD
D000818:Animals; D001483:Base Sequence; D016825:Chlamydomonas reinhardtii / Q000235:genetics*; D002736:Chloroplasts / Q000502:physiology*; D004262:DNA Restriction Enzymes; D015252:Deoxyribonucleases, Type II Site-Specific / Q000096:biosynthesis / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D014644:Genetic Variation; D007438:Introns / Q000235:genetics*; D008969:Molecular Sequence Data; D012338:RNA, Ribosomal, 23S / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis; D013379:Substrate Specificity
D012338:RNA, Ribosomal, 23S; D011994:Recombinant Proteins; D004262:DNA Restriction Enzymes; C048901:endodeoxyribonuclease CreI; D015252:Deoxyribonucleases, Type II Site-Specific