Transposition of a group II intron.
ABSTRACT Among mobile genetic elements, self-splicing introns are of particular interest. They belong to either group I or group II depending on their three-dimensional structure. Homing, the systematic intron invasion of an intronless gene when it encounters its homologous intron-bearing allele, is the only means for intron mobility so far demonstrated. It depends on the activity of the intron-encoded protein and is very specific for the acceptor site. Intron transposition, the transfer of an intron to a novel site, predicted on the basis of phylogenetic studies and in vitro reverse-splicing experiments, has been proposed to be responsible for evolutionary intron spreading. Here we present results from polymerase chain reaction experiments consistent with transposition of a group II intron. This event is proposed to account for the site-specific deletion in the mitochondrial chromosome of the fungus Podospora anserina that is associated with the premature death syndrome and might also be involved in the senescence process affecting this species.
Article: Mobile group II introns.[show abstract] [hide abstract]
ABSTRACT: Mobile group II introns, found in bacterial and organellar genomes, are both catalytic RNAs and retrotransposable elements. They use an extraordinary mobility mechanism in which the excised intron RNA reverse splices directly into a DNA target site and is then reverse transcribed by the intron-encoded protein. After DNA insertion, the introns remove themselves by protein-assisted, autocatalytic RNA splicing, thereby minimizing host damage. Here we discuss the experimental basis for our current understanding of group II intron mobility mechanisms, beginning with genetic observations in yeast mitochondria, and culminating with a detailed understanding of molecular mechanisms shared by organellar and bacterial group II introns. We also discuss recently discovered links between group II intron mobility and DNA replication, new insights into group II intron evolution arising from bacterial genome sequencing, and the evolutionary relationship between group II introns and both eukaryotic spliceosomal introns and non-LTR-retrotransposons. Finally, we describe the development of mobile group II introns into gene-targeting vectors, "targetrons," which have programmable target specificity.Annual Review of Genetics 02/2004; 38:1-35. · 22.23 Impact Factor
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ABSTRACT: Group II intron homing in yeast mitochondria is initiated at active target sites by activities of intron-encoded ribonucleoprotein (RNP) particles, but is completed by competing recombination and repair mechanisms. Intron aI1 transposes in haploid cells at low frequency to target sites in mtDNA that resemble the exon 1-exon 2 (E1/E2) homing site. This study investigates a system in which aI1 can transpose in crosses (i.e., in trans). Surprisingly, replacing an inefficient transposition site with an active E1/E2 site supports <1% transposition of aI1. Instead, the ectopic site was mainly converted to the related sequence in donor mtDNA in a process we call "abortive transposition." Efficient abortive events depend on sequences in both E1 and E2, suggesting that most events result from cleavage of the target site by the intron RNP particles, gapping, and recombinational repair using homologous sequences in donor mtDNA. A donor strain that lacks RT activity carries out little abortive transposition, indicating that cDNA synthesis actually promotes abortive events. We also infer that some intermediates abort by ejecting the intron RNA from the DNA target by forward splicing. These experiments provide new insights to group II intron transposition and homing mechanisms in yeast mitochondria.Genetics 09/2004; 168(1):77-87. · 4.01 Impact Factor
Article: Compilation and analysis of group II intron insertions in bacterial genomes: evidence for retroelement behavior.[show abstract] [hide abstract]
ABSTRACT: Group II introns are novel genetic elements that have properties of both catalytic RNAs and retroelements. Initially identified in organellar genomes of plants and lower eukaryotes, group II introns are now being discovered in increasing numbers in bacterial genomes. Few of the newly sequenced bacterial introns are correctly identified or annotated by those who sequenced them. Here we have compiled and thoroughly analyzed group II introns and their fragments in bacterial DNA sequences reported to GenBank. Intron distribution in bacterial genomes differs markedly from the distribution in organellar genomes. Bacterial introns are not inserted into conserved genes, are often inserted outside of genes altogether and are frequently fragmented, suggesting a high rate of intron gain and loss. Some introns have multiple natural homing sites while others insert after transcriptional terminators. All bacterial group II introns identified to date encode reverse transcriptase open reading frames and are either active retroelements or derivatives of retroelements. Together, these observations suggest that group II introns in bacteria behave primarily as retroelements rather than as introns, and that the strategy for group II intron survival in bacteria is fundamentally different from intron survival in organelles.Nucleic Acids Research 04/2002; 30(5):1091-102. · 8.03 Impact Factor