Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 447805
Title CRISPR-Cas systems preferentially target the leading regions of MOBF conjugative plasmids
Author(s) Westra, E.R.; Staals, R.H.J.; Gort, G.; Høgh, S.; Neumann, S.; Cruz, F.; Fineran, P.C.; Brouns, S.J.J.
Source RNA Biology 10 (2013)5. - ISSN 1547-6286 - p. 749 - 761.
DOI https://doi.org/10.4161/rna.24202
Department(s) Microbiological Laboratory
Biometris (WU MAT)
VLAG
Publication type Refereed Article in a scientific journal
Publication year 2013
Keyword(s) horizontal gene-transfer - protein h-ns - adaptive immune-systems - escherichia-coli - streptococcus-thermophilus - host-range - in-vitro - microbial communities - acquired-resistance - antiviral defense
Abstract Most prokaryotes contain CRISPR-Cas immune systems that provide protection against mobile genetic elements. We have focused on the ability of CRISPR-Cas to block plasmid conjugation, and analyzed the position of target sequences (protospacers) on conjugative plasmids. The analysis reveals that protospacers are non-uniformly distributed over plasmid regions in a pattern that is determined by the plasmid's mobilization type (MOB). While MOBP plasmids are most frequently targeted in the region entering the recipient cell last (lagging region), MOBF plasmids are mostly targeted in the region entering the recipient cell first (leading region). To explain this protospacer distribution bias, we propose two mutually non-exclusive hypotheses: (1) spacers are acquired more frequently from either the leading or lagging region depending on the MOB type (2) CRISPR-interference is more efficient when spacers target these preferred regions. To test the latter hypothesis, we analyzed Type I-E CRISPR-interference against MOBF prototype plasmid F in Escherichia coli. Our results show that plasmid conjugation is effectively inhibited, but the level of immunity is not affected by targeting the plasmid in the leading or lagging region. Moreover, CRISPR-immunity levels do not depend on whether the incoming single-stranded plasmid DNA, or the DNA strand synthesized in the recipient is targeted. Our findings indicate that single-stranded DNA may not be a target for Type I-E CRISPR-Cas systems, and suggest that the protospacer distribution bias might be due to spacer acquisition preferences
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