In vitroself-splicing reactions of the chloroplast group I intron Cr.LSU fromChlamydomonas reinhardtiiand in vivo manipulation via gene-replacement
Open Access
- 1 January 1991
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 19 (23) , 6611-6618
- https://doi.org/10.1093/nar/19.23.6611
Abstract
The group I intron from the chloroplast rRNA large subunit of Chlamydomonas reinhardtii (Cr.LSU) undergoes autocatalytic splicing in vitro. Cr.LSU displays a range of reactions typical of other group I introns. Under optimal conditions, the 5′ cleavage step proceeds rapidly, but the exon-ligation step is relatively slow, and no pH dependent hydrolysis of the 3′ splice site occurs. A requirement for high temperature and high [Mg2+[ suggests involvement of additional splicing factors in vivo. The positions of three cyclization sites of the free intron have been mapped; two of these sites represent reactions analogous to 5′-splice site cleavage, whereas the third is an example of G-exchange. Cr.LSU contains an open reading frame (ORF) potentially encoding an 163 amino acid polypeptide. ORF function has been investigated by using chloroplast gene replacement via particle bombardment. We have shown that the ORF can be deleted from Cr.LSU without affecting splicing in vivo and it thus does not encode an essential splicing factor.Keywords
This publication has 41 references indexed in Scilit:
- Group I Intron Self-Splicing with Adenosine: Evidence for a Single Nucleoside-Binding SiteScience, 1991
- Effects of mutations of the bulged nucleotide in the conserved P7 pairing element of the phage T4 td intron on ribozyme functionBiochemistry, 1991
- Six group I introns and three internal transcribed spacers in the chloroplast large subunit ribosomal RNA gene of the green alga Chlamydomonas eugametosJournal of Molecular Biology, 1991
- Selection of small molecules by the Tetrahymena catalytic centerNucleic Acids Research, 1991
- The rate and specificity of a group I ribozyme are inversely affected by choice of monovalent saltNucleic Acids Research, 1991
- 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 intronCurrent Genetics, 1991
- Selection of circularization sites in a group I IVS RNA requires multiple alignments of an internal template-like sequenceCell, 1987
- Reactions of the intervening sequence of the Tetrahymena ribosomal ribonucleic acid precursor: pH dependence of cyclization and site-specific hydrolysisBiochemistry, 1985
- Splicing of large ribosomal precursor RNA and processing of intron RNA in yeast mitochondriaCell, 1984
- Autocatalytic cyclization of an excised intervening sequence RNA is a cleavage–ligation reactionNature, 1983