Target Motifs Affecting Natural Immunity by a Constitutive CRISPR-Cas System in Escherichia coli

被引：47

作者：

Almendros, Cristobal ^{[1
]}

Guzman, Noemi M. ^{[1
]}

Diez-Villasenor, Cesar ^{[1
]}

Garcia-Martinez, Jesus ^{[1
]}

Mojica, Francisco J. M. ^{[1
]}

机构：

[1] Univ Alicante, Fac Ciencias, Dept Fisiol Genet & Microbiol, E-03080 Alicante, Spain

来源：

PLOS ONE | 2012年 / 7卷 / 11期

关键词：

RNA; DNA; SEQUENCE; BACTERIOPHAGE; TRANSCRIPTION; RESISTANCE; REPEATS; DEFENSE; IDENTIFICATION; INTERFERENCE;

D O I：

10.1371/journal.pone.0050797

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR associated (cas) genes conform the CRISPR-Cas systems of various bacteria and archaea and produce degradation of invading nucleic acids containing sequences (protospacers) that are complementary to repeat intervening spacers. It has been demonstrated that the base sequence identity of a protospacer with the cognate spacer and the presence of a protospacer adjacent motif (PAM) influence CRISPR-mediated interference efficiency. By using an original transformation assay with plasmids targeted by a resident spacer here we show that natural CRISPR-mediated immunity against invading DNA occurs in wild type Escherichia coli. Unexpectedly, the strongest activity is observed with protospacer adjoining nucleotides (interference motifs) that differ from the PAM both in sequence and location. Hence, our results document for the first time native CRISPR activity in E. coli and demonstrate that positions next to the PAM in invading DNA influence their recognition and degradation by these prokaryotic immune systems.

引用

页数：7

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