Homing of a bacterial group II intron with an intron‐encoded protein lacking a recognizable endonuclease domain
Open Access
- 1 March 2000
- journal article
- research article
- Published by Wiley in Molecular Microbiology
- Vol. 35 (6) , 1405-1412
- https://doi.org/10.1046/j.1365-2958.2000.01804.x
Abstract
RmInt1 is a functional group II intron found in Sinorhizobium meliloti where it interrupts a group of IS elements of the IS630‐Tc1 family. In contrast to many other group II introns, the intron‐encoded protein (IEP) of RmInt1 lacks the characteristic conserved part of the Zn domain associated with the IEP endonuclease activity. Nevertheless, in this study, we show that RmInt1 is capable of inserting into a vector containing the DNA spanning the RmInt1 target site from the genome of S. meliloti. Efficient homing was also observed in the absence of homologous recombination (RecA− strains). In addition, it is shown that RmInt1 is able to move to its target in a heterologous host (S. medicae). Homing of RmInt1 occurs very efficiently upon DNA target uptake (conjugation/electroporation) by the host cell resulting in a proportion of invaded target of 11–30%. Afterwards, the remaining intronless target DNA is protected from intron invasion.Keywords
This publication has 44 references indexed in Scilit:
- Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicumPublished by Elsevier ,2003
- RNA and Protein Catalysis in Group II Intron Splicing and Mobility Reactions Using Purified ComponentsBiochemistry, 1999
- Group II intron reverse transcriptase in yeast mitochondria. Stabilization and regulation of reverse transcriptase activity by the intron RNAJournal of Molecular Biology, 1999
- Group II intron mobility in yeast mitochondria: target DNA-primed reverse transcription activity of ai1 and reverse splicing into DNA transposition sites in vitroJournal of Molecular Biology, 1998
- Group II intron endonucleases use both RNA and protein subunits for recognition of specific sequences in double-stranded DNAThe EMBO Journal, 1997
- Gapped BLAST and PSI-BLAST: a new generation of protein database search programsNucleic Acids Research, 1997
- Efficient integration of an intron RNA into double-stranded DNA by reverse splicingNature, 1996
- Group II self-splicing introns in bacteriaNature, 1993
- Reverse transcriptase activity associated with maturase-encoding group II introns in yeast mitochondriaCell, 1993
- Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti.Proceedings of the National Academy of Sciences, 1980