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Characterisation of a novel homing endonuclease and screening of intervening sequences in closely related Bacillus thuringiensis nrdIEF genes
Stockholm University, Faculty of Science, Department of Molecular Biology and Functional Genomics.
Article in journal (Refereed) Submitted
URN: urn:nbn:se:su:diva-24434OAI: diva2:197525
Part of urn:nbn:se:su:diva-7042Available from: 2007-08-30 Created: 2007-08-30Bibliographically approved
In thesis
1. Homing Endonucleases and Horizontal Gene Transfer in Bacteria and Bacteriophages
Open this publication in new window or tab >>Homing Endonucleases and Horizontal Gene Transfer in Bacteria and Bacteriophages
2007 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Homing endonuclease genes (HEGs) are selfish genetic elements that mediate their own super-Mendelian inheritance. This is mediated by the homing endonuclease cleavage of a HEG- allele followed by recombination-repair with a HEG+ allele.

The majority of the HEGs are encoded in intervening sequences (IVSs). The insertion of the IVS interrupts the endonuclease recognition site, making the genome with the IVS resistant to further cleavage by homing endonucleases with specificity for that particular sequence, but susceptible for homing endonucleases with a target not affected by the IVS insert. In 39 studied strains of the Bacillus cereus group, 28 IVSs were found in the nrdIEF operon. Phylogenetic studies of these sequences showed a scattered distribution of the IVSs, indicating a frequent horizontal gene transfer and that there might be competition between the different IVSs in the nrdIEF operon in the Bacillaceae family. One novel group I intron was shown to encode a functional homing endonuclease with a GIY-(X)8-YIG motif, expanding the family motif to GIY-(X)8-11-YIG. Interestingly, by studying the known insertion sites for IVSs in the ribonuclotide reductase genes, we show that the majority of the insertions are at conserved motifs, indicating that conservation is important for IVS survival.

Most freestanding HEGs in bacteriophage T4 cleave both HEG+ and HEG- alleles, possibly providing a competitive advantage for the host organism when two phages infect the same bacterium. Two novel freestanding HEGs replace two putative HEGs in T4 in some T-even-like phages. The characterisation of these HEGs showed that both cleave double stranded DNA. SegH was shown to promote homing of its gene. Hef showed no homing, possibly due to general exclusion of other phages. The mobE putative HEG was shown to be homing proficient and showed strong general DNA degradation when expressed in Escherichia coli.

Place, publisher, year, edition, pages
Stockholm: Institutionen för molekylärbiologi och funktionsgenomik, 2007. 70 p.
dsDNA cleavage, ribonucleotide reductase, GIY-YIG motif
National Category
Biochemistry and Molecular Biology
Research subject
Molecular Biology
urn:nbn:se:su:diva-7042 (URN)978-91-7155-477-2 (ISBN)
Public defence
2007-09-21, Nordenskiöldsalen, Geovetenskapens hus, Svante Arrhenius väg 8 C, Stockholm, 10:00
Available from: 2007-08-30 Created: 2007-08-30Bibliographically approved

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