Cleavage and recognition pattern of a double-strand-specific endonuclease (I-CreI) encoded by the chloroplast 23S rRNA intron of Chlamydomonas reinhardtii

Thompson, Andrew J.; Yuan, Xiaoqin; Kudlicki, Wiesław; Herrin, David L.

doi:10.1016/0378-1119(92)90278-w

Cited by 67 publications

(49 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The yeast mitochondrial enzymes I-SceI and I-SceII each recognize 18-bp sequences (7,44,54), while the algal chloroplast endonuclease I-CreI requires a 24-bp sequence for cleavage (51). I-TevI has a relatively large recognition sequence of ca.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Interaction of the intron-encoded mobility endonuclease I-PpoI with its target site.

Ellison¹,

Vogt²

1993

Mol. Cell. Biol.

View full text Add to dashboard Cite

Endonucleases encoded by mobile group I introns are highly specific DNases that induce a double-strand break near the site to which the intron moves. I-PpoI from the acellular slime mold Physarum polycephalum mediates the mobility of intron 3 (Pp LSU 3) in the extrachromosomal nuclear ribosomal DNA of this organism. We showed previously that cleavage by I-PpoI creates a four-base staggered cut near the point of intron insertion. We have now characterized several further properties of the endonuclease. As determined by deletion analysis, the minimal target site recognized by I-PpoI was a sequence of 13 to 15 bp spanning the cleavage site. The purified protein behaved as a globular dimer in sedimentation and gel filtration. In gel mobility shift assays in the presence of EDTA, I-PpoI formed a stable and specific complex with DNA, dissociating with a half-life of 45 min. By footprinting and interference assays with methidiumpropyl-EDTAiron(II), I-PpoI contacted a 22-to 24-bp stretch of DNA. The endonuclease protected most of the purines found in both the major and minor grooves of the DNA helix from modification by dimethyl sulfate (DMS). However, the reactivity to DMS was enhanced at some purines, suggesting that binding leads to a conformational change in the DNA. The pattern of DMS protection differed fundamentally in the two partially symmetrical halves of the recognition sequence.More than 100 group I introns, which are defined by a common RNA secondary structure and mechanism of splicing, have been identified in the organellar genomes of lower eucaryotes (for reviews, see references 4 and 29). Several of these introns also have been found in bacteria, bacteriophage, and extrachromosomal nuclear ribosomal DNA (rDNA). The RNAs of many group I introns are capable of self-splicing in vitro. Some group I introns behave as mobile elements, as first described for the optional omega intron (Sc LSU 1) of Saccharomyces cerevisiae (9). Additional mobile group I introns have been found in yeast mitochondria (32,45,53), bacteriophage (42), Chlamydomonas chloroplasts (11,23), Chlamydomonas mitochondria (8, 26), and nuclear rDNA of slime molds (16,36). In each of these cases, the mobile intron is copied into an intron-allele of the gene in which it resides, converting it to intron+ by gene conversion initiated at a double-strand break near the site of insertion. This process, known as intron homing (10), is mediated by an endonuclease encoded within the intron, which recognizes and cleaves the intron-DNA within a few nucleotides of the insertion site (for reviews, see references 22 and 33). In three cases, the procaryotic enzymes I-TevI, I-TevII, and I-TevIII, the DNA cut is somewhat more removed, 13 to 26 bp from the point of intron insertion (for a review, see reference 33). In any case, the homology between the intron-allele and the DNA sequences flanking the intron in the intron+ allele allows the break to be repaired via a gene conversion event which uses the intron+ allele as a template. Since intron insertion render...

show abstract

Section: Discussionmentioning

confidence: 99%

“…The specificity of a few intronencoded endonucleases has been investigated experimentally. The yeast mitochondrial enzymes I-SceI and I-SceII recognize 18-bp sequences (7,54), and the algal chloroplast protein I-CreI is reported to require a 24-bp sequence for cleavage (51). The phage enzyme I-TevI recognizes ca.…”

mentioning

confidence: 99%

Interaction of the intron-encoded mobility endonuclease I-PpoI with its target site.

Ellison¹,

Vogt²

1993

Mol. Cell. Biol.

View full text Add to dashboard Cite

show abstract

“…We speculate that such doublestrand cuts were generated by the endonuclease encoded by the invading ORFs, leading to the integration of these ORFs and the production of mobile introns in ancestral Chlamydomonas chloroplast rmL genes. By analyzing the intron sequences in the immediate vicinity of the ORFs specifying I-CeuI and I-CreI in the C. eugametos and Chlamydomonas reinhardtii (Durrenberger andRochaix, 1991, 1993 ;Rochaix et al, 1985;Thompson et al, 1992) chloroplast rmL genes, we found that sequences immediately preceding and following these ORFs display significant similarity to exon sequences flanking the insertion sites of the respective introns. As shown in Fig.…”

Section: Role Of Homing Endonucleases In the Evolution Of Chloroplastmentioning

confidence: 93%

The I‐Ceu I endonuclease: purification and potential role in the evolution of Chlamydomonas group I introns

Marshall

Davis

Lemieux

1994

European Journal of Biochemistry

View full text Add to dashboard Cite

During genetic crosses between the interfertile green algae Chlamydomonas eugametos and Chlamydomonas moewusii, the I‐Ceul endonuclease encoded by the fifth group I intron (CeLSU · 5) in the C. eugametos chloroplast large subunit rRNA gene mediates the mobility of this intron by introducing a double‐strand break near the insertion site of the intron in the corresponding C. moewusii intronless allele. To characterize the biochemical properties of this endonuclease, we have purified I‐CeuI and determined the optimal reaction conditions for cleavage. I‐CeuI activity is maximal at 50°C, pH 10.0, 2.5 mM MgCl2 and in the absence of NaCl. Unlike the well‐characterized I‐SceI endonuclease, I‐CeuI remains stable following preincubation in the absence of substrate. We discuss the role that homing endonucleases may have played in the evolution of Chlamydomonas chloroplast group I introns.

show abstract

“…Meganucleases are homing endonucleases derived from I-CreI [7] and I-CeuI [8] and operate as homodimers that recognize DNA sequences with palindromic sequences. Variants containing two motifs, such as I-SceI, act as monomers consisting of a pair of nuclease domains with symmetry-independent targeting [9].…”

Section: Toolsmentioning

confidence: 99%

Genome-Edited T Cell Therapies

Delhove

Qasim

2017

Curr Stem Cell Rep

View full text Add to dashboard Cite

Purpose of ReviewAlternative approaches to conventional drug-based cancer treatments have seen T cell therapies deployed more widely over the last decade. This is largely due to their ability to target and kill specific cell types based on receptor recognition. Introduction of recombinant T cell receptors (TCRs) using viral vectors and HLA-independent T cell therapies using chimeric antigen receptors (CARs) are discussed. This article reviews the tools used for genome editing, with particular emphasis on the applications of site-specific DNA nuclease mediated editing for T cell therapies.Recent FindingsGenetic engineering of T cells using TCRs and CARs with redirected antigen-targeting specificity has resulted in clinical success of several immunotherapies. In conjunction, the application of genome editing technologies has resulted in the generation of HLA-independent universal T cells for allogeneic transplantation, improved T cell sustainability through knockout of the checkpoint inhibitor, programmed cell death protein-1 (PD-1), and has shown efficacy as an antiviral therapy through direct targeting of viral genomic sequences and entry receptors.SummaryThe combined use of engineered antigen-targeting moieties and innovative genome editing technologies have recently shown success in a small number of clinical trials targeting HIV and hematological malignancies and are now being incorporated into existing strategies for other immunotherapies.

show abstract

Cleavage and recognition pattern of a double-strand-specific endonuclease (I-CreI) encoded by the chloroplast 23S rRNA intron of Chlamydomonas reinhardtii

Cited by 67 publications

References 18 publications

Interaction of the intron-encoded mobility endonuclease I-PpoI with its target site.

Interaction of the intron-encoded mobility endonuclease I-PpoI with its target site.

The I‐Ceu I endonuclease: purification and potential role in the evolution of Chlamydomonas group I introns

Genome-Edited T Cell Therapies

Contact Info

Product

Resources

About