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Caspase, a family of cysteine proteases, serve as effectors in apoptosis. The ced3 gene in C.elegans was first identified to be involved in apoptosis. This gene encodes the ced-3 caspase that is similar to the interleukin-1-beta converting enzyme or ICE in mammals. In addition to apoptosis, caspases also function in the inflammatory response. Inflammatory caspases are essential in activating pro-inflammatory cytokines that recruit immune cells and block the replication of pathogens inside...
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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
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CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
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At the transition from prophase to metaphase, there is a reduction in cohesion along the chromosomal arms, resulting in the resolution of sister chromatids. However, residual cohesin connections remain to hold the sister chromatids together until the transition from metaphase to anaphase. The residual connection prevents any premature separation of sister chromatids, blocking the risks of aneuploidy within the daughter cells.
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Substrate Generation for Endonucleases of CRISPR/Cas Systems
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Disintegration promotes protospacer integration by the Cas1-Cas2 complex.

Chien-Hui Ma1, Kamyab Javanmardi1, Ilya J Finkelstein1,2

  • 1Department of Molecular Biosciences and Institute of Cell and Molecular Biology, University of Texas at Austin, Austin, United States.

Elife
|August 26, 2021
PubMed
Summary

Disintegration, a reversal of DNA integration, is not abortive but a key step in CRISPR immunity. This process, involving the Cas1-Cas2 complex, facilitates protospacer insertion and bacterial defense.

Keywords:
CRISPRE. coliadaptationbiochemistrychemical biologytransposition

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Area of Science:

  • Molecular Biology
  • Bacterial Immunity
  • DNA Transposition

Background:

  • Disintegration, the reversal of DNA integration, is traditionally viewed as an abortive reaction.
  • The Streptococcus pyogenes Cas1-Cas2 complex mediates protospacer insertion, a process analogous to DNA transposition.

Purpose of the Study:

  • To biochemically investigate the mechanism of protospacer insertion by the Cas1-Cas2 complex.
  • To challenge the view of disintegration as solely an abortive off-pathway reaction.

Main Methods:

  • Biochemical investigation of protospacer insertion mechanism.
  • Analysis of integration outcomes in supercoiled and linear target DNA sites.

Main Results:

  • Disintegration is the predominant outcome for one-ended insertions that fail to complete integration in supercoiled targets.
  • In linear targets, one-ended insertions are more frequent than complete insertions, often accompanied by disintegration of the first insertion.
  • One-ended integration intermediates can mature into complete spacer insertions via DNA repair pathways.

Conclusions:

  • Disintegration is functionally important in the adaptive phase of CRISPR-mediated bacterial immunity.
  • Disintegration-promoted integration may play a role in other transposition reactions.
  • The Cas1-Cas2 complex's mechanism highlights the significance of disintegration in biological processes.