A group I intron in the chloroplast large subunit rRNA gene of Chlamydomonas eugametos encodes a double-strand endonuclease that cleaves the homing site of this intron.
During interspecific crosses between Chlamydomonas eugametos and Chlamydomonas moewusii, an optional group I intron of 955 base pairs (CeLSU.5) in the C. eugametos chloroplast large subunit rRNA gene undergoes a duplicative transposition event which is associated with frequent co-conversion of flanking cpDNA sequences. In the present study, we show that the basic protein of 218 amino acids encoded by CeLSU.5 could mediate the phenomenon of intron transposition, also called intron homing. We overexpressed the ORF specifying this protein in E. coli using expression vectors that contain a C. moewusii cpDNA sequence encompassing the intron homing site. The expression product was found to exhibit a double-strand DNA endonuclease activity that is specific for the homing site. This activity was detected in vivo by self-linearization of the expression plasmids.
Gauthier
A
A
Départment de biochimie, Faculté des Sciences et de Génie, Université Laval, Québec, Canada.
Turmel
M
M
Lemieux
C
C
D000595:Amino Acid Sequence; D001483:Base Sequence; D002696:Chlamydomonas / Q000201:enzymology / Q000235:genetics*; D002736:Chloroplasts; D004275:DNA, Ribosomal / Q000235:genetics* / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000235:genetics* / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000235:genetics* / Q000378:metabolism; D005822:Genetic Vectors; D007438:Introns; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D012335:RNA, Ribosomal / Q000235:genetics*
D004275:DNA, Ribosomal; D012335:RNA, Ribosomal; D004706:Endodeoxyribonucleases; C052380:endodeoxyribonuclease NcoI; D015252:Deoxyribonucleases, Type II Site-Specific