Repeat Size Determination by Two Molecular Rulers in the Type I-E CRISPR Array.
Prokaryotic adaptive immune systems are composed of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) proteins. These systems adapt to new threats by integrating short nucleic acids, termed spacers, into the CRISPR array. The functional motifs in the repeat and the mechanism by which a constant repeat size is maintained are still elusive. Here, through a series of mutations within the repeat of the CRISPR-Cas type I-E, we identify motifs that are crucial for adaptation and show that they serve as anchor sites for two molecular rulers determining the size of the new repeat. Adaptation products from various repeat mutants support a model in which two motifs in the repeat bind to two different sites in the adaptation complex that are 8 and 16 bp away from the active site. This model significantly extends our understanding of the adaptation process and broadens the scope of its applications.
Department of Clinical Microbiology and Immunology, Sackler School of Medicine, Tel Aviv University, 69978 Tel Aviv, Israel.
Moran G
MG
Goren
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot 76100, Israel.
Shany
S
Doron
Department of Clinical Microbiology and Immunology, Sackler School of Medicine, Tel Aviv University, 69978 Tel Aviv, Israel.
Rea
R
Globus
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot 76100, Israel.
Gil
G
Amitai
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot 76100, Israel. Electronic address: rotem.sorek@weizmann.ac.il.
Rotem
R
Sorek
Department of Clinical Microbiology and Immunology, Sackler School of Medicine, Tel Aviv University, 69978 Tel Aviv, Israel. Electronic address: ehudq@post.tau.ac.il.
Udi
U
Qimron
D001483:Base Sequence; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005819:Genetic Markers; D008957:Models, Genetic; D059372:Nucleotide Motifs / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*; D015203:Reproducibility of Results
D005819:Genetic Markers
adaptation; anchor site; elongated repeat; shortened repeat; spacer integration