scholarly journals The gRAMP CRISPR-Cas effector is an RNA endonuclease complexed with a caspase-like peptidase

Science ◽  
2021 ◽  
Vol 373 (6561) ◽  
pp. 1349-1353
Author(s):  
Sam P. B. van Beljouw ◽  
Anna C. Haagsma ◽  
Alicia Rodríguez-Molina ◽  
Daan F. van den Berg ◽  
Jochem N. A. Vink ◽  
...  
Keyword(s):  
Rna Endonuclease

High-resolution crystal structure reveals a HEPN domain at the C-terminal region of S. cerevisiae RNA endonuclease Swt1

2014 ◽  
Vol 453 (4) ◽  
pp. 826-832 ◽  
Author(s):  
Shuxia Peng ◽  
Ke Zhou ◽  
Wenjia Wang ◽  
Zengqiang Gao ◽  
Yuhui Dong ◽  
...  
Keyword(s):  
Crystal Structure ◽  
High Resolution ◽  
Terminal Region ◽  
Rna Endonuclease

scholarly journals PPR-SMR protein SOT1 has RNA endonuclease activity

2017 ◽  
Vol 114 (8) ◽  
pp. E1554-E1563 ◽  
Author(s):  
Wen Zhou ◽  
Qingtao Lu ◽  
Qingwei Li ◽  
Lei Wang ◽  
Shunhua Ding ◽  
...  
Keyword(s):  
Endonuclease Activity ◽  
Pentatricopeptide Repeat ◽  
Rna Sequence ◽  
Ppr Protein ◽  
Specific Manner ◽  
Rna Endonuclease

Numerous attempts have been made to identify and engineer sequence-specific RNA endonucleases, as these would allow for efficient RNA manipulation. However, no natural RNA endonuclease that recognizes RNA in a sequence-specific manner has been described to date. Here, we report that SUPPRESSOR OF THYLAKOID FORMATION 1 (SOT1), anArabidopsispentatricopeptide repeat (PPR) protein with a small MutS-related (SMR) domain, has RNA endonuclease activity. We show that the SMR moiety of SOT1 performs the endonucleolytic maturation of 23S and 4.5S rRNA through the PPR domain, specifically recognizing a 13-nucleotide RNA sequence in the 5′ end of the chloroplast 23S–4.5S rRNA precursor. In addition, we successfully engineered the SOT1 protein with altered PPR motifs to recognize and cleave a predicted RNA substrate. Our findings point to SOT1 as an exciting tool for RNA manipulation.

scholarly journals Improved, two-stage protein expression and purification via autoinduction of both autolysis and auto DNA/RNA hydrolysis conferred by phage lysozyme and DNA/RNA endonuclease

2020 ◽  
Author(s):  
Romel Menacho-Melgar ◽  
Eirik A. Moreb ◽  
John P. Efromson ◽  
Michael D. Lynch
Keyword(s):  
Protein Expression ◽  
Membrane Integrity ◽  
Heterologous Protein Expression ◽  
List Type ◽  
Two Stage ◽  
Microtiter Plates ◽  
Thaw Cycle ◽  
Protein Expression And Purification ◽  
Phosphate Depletion ◽  
Rna Endonuclease

AbstractWe report improved release of recombinant proteins in E. coli, which relies on combined cellular autolysis and DNA/RNA autohydrolysis, conferred by the tightly controlled autoinduction of both phage lysozyme and the non specific DNA/RNA endonuclease from S. marcescens. Autoinduction occurs in a two-stage process wherein heterologous protein expression and autolysis enzymes are induced upon entry into stationary phase by phosphate depletion. Cytoplasmic lysozyme and periplasmic endonuclease are kept from inducing lysis until membrane integrity is disrupted. Post cell harvest, the addition of detergent (0.1% Triton-X100) and a single 30 minutes freezer thaw cycle results in > 90% release of protein (GFP). This cellular lysis is accompanied by complete oligonucleotide hydrolysis. The approach has been validated for shake flask cultures, high throughput cultivation in microtiter plates and larger scale stirred-tank bioreactors. This tightly controlled system enables robust growth and resistance to lysis in routine media when cells are propagated and autolysis/hydrolysis genes are only induced upon phosphate depletion.HighlightsAutoinduction of both cell lysis and nucleotide hydrolysis>90 % lysis and DNA degradationStrains are stable to lysis in the absence of phosphate depletion.

scholarly journals RISC is a 5' phosphomonoester-producing RNA endonuclease

2004 ◽  
Vol 18 (9) ◽  
pp. 975-980 ◽  
Author(s):  
J. Martinez
Keyword(s):  
Rna Endonuclease

scholarly journals In vivo analysis of trypanosome mitochondrial RNA function by artificial site-specific RNA endonuclease-mediated knockdown

RNA ◽  
2015 ◽  
Vol 21 (10) ◽  
pp. 1781-1789 ◽  
Author(s):  
Anthony J. Szempruch ◽  
Rajarshi Choudhury ◽  
Zefeng Wang ◽  
Stephen L. Hajduk
Keyword(s):  
Mitochondrial Rna ◽  
Site Specific ◽  
In Vivo Analysis ◽  
Rna Endonuclease

scholarly journals The Integrator complex terminates promoter-proximal transcription at protein-coding genes

2019 ◽  
Author(s):  
Nathan D. Elrod ◽  
Telmo Henriques ◽  
Kai-Lieh Huang ◽  
Deirdre C. Tatomer ◽  
Jeremy E. Wilusz ◽  
...  
Keyword(s):  
Rna Polymerase Ii ◽  
List Type ◽  
Protein Coding ◽  
Pol Ii ◽  
Bacterial Transcription ◽  
Number Of Genes ◽  
Nascent Rna ◽  
Template Dna ◽  
Productive Elongation ◽  
Rna Endonuclease

SUMMARYThe transition of RNA polymerase II (Pol II) from initiation to productive elongation is a central, regulated step in metazoan gene expression. At many genes, Pol II pauses stably in early elongation, remaining engaged with the 25-60 nucleotide-long nascent RNA for many minutes while awaiting signals for release into the gene body. However, a number of genes display highly unstable promoter Pol II, suggesting that paused polymerase might dissociate from template DNA at these promoters and release a short, non-productive mRNA. Here, we report that paused Pol II can be actively destabilized by the Integrator complex. Specifically, Integrator utilizes its RNA endonuclease activity to cleave nascent RNA and drive termination of paused Pol II. These findings uncover a previously unappreciated mechanism of metazoan gene repression, akin to bacterial transcription attenuation, wherein promoter-proximal Pol II is prevented from entering productive elongation through factor-regulated termination.HighlightsThe Integrator complex inhibits transcription elongation at ∼15% of mRNA genesIntegrator targets promoter-proximally paused Pol II for terminationThe RNA endonuclease of Integrator subunit 11 is critical for gene attenuationIntegrator-repressed genes are enriched in signaling and growth-responsive pathways

Tools for Investigation of the RNA Endonuclease Activity of Mammalian Argonaute2 Protein

2012 ◽  
Vol 84 (5) ◽  
pp. 2492-2497 ◽  
Author(s):  
Nana Yang ◽  
Ya Cao ◽  
Ping Han ◽  
Xuejun Zhu ◽  
Lizhou Sun ◽  
...  
Keyword(s):  
Endonuclease Activity ◽  
Rna Endonuclease
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