Ryan N. Jackson, Blake Wiedenheft  Molecular Cell 

Slides:



Advertisements
Similar presentations
Volume 28, Issue 2, Pages (October 2007)
Advertisements

Box H/ACA Small Ribonucleoproteins
Presented By: Diana Marquez
by Prarthana Mohanraju, Kira S
A CRISPR View of Cleavage
Expanding the Biologist’s Toolkit with CRISPR-Cas9
CRISPR-Cas Systems: Prokaryotes Upgrade to Adaptive Immunity
Volume 56, Issue 4, Pages (November 2014)
Intrinsic Cellular Defenses against Human Immunodeficiency Viruses
Volume 62, Issue 2, Pages (April 2016)
Structural Basis of DNA Recognition by p53 Tetramers
Rewiring Cas9 to Target New PAM Sequences
Ubiquitination Accomplished: E1 and E2 Enzymes Were Not Necessary
Chaoyou Xue, Natalie R. Whitis, Dipali G. Sashital  Molecular Cell 
Archaeal CRISPR-based immune systems: exchangeable functional modules
Volume 60, Issue 3, Pages (November 2015)
Targeting HSP70 for Cancer Therapy
Volume 56, Issue 1, Pages (October 2014)
Volume 26, Issue 2, Pages (April 2007)
HOPX: The Unusual Homeodomain-Containing Protein
Volume 45, Issue 3, Pages (February 2012)
Takuo Osawa, Hideko Inanaga, Chikara Sato, Tomoyuki Numata 
Daan C. Swarts, John van der Oost, Martin Jinek  Molecular Cell 
Volume 28, Issue 2, Pages (October 2007)
CRISPR-Cas Systems: Prokaryotes Upgrade to Adaptive Immunity
Volume 40, Issue 4, Pages (November 2010)
Structure of the Replicating Complex of a Pol α Family DNA Polymerase
Addison V. Wright, James K. Nuñez, Jennifer A. Doudna  Cell 
Volume 34, Issue 4, Pages (May 2009)
Structure of the Endonuclease Domain of MutL: Unlicensed to Cut
Alpers-Huttenlocher Syndrome
Single-Stranded DNA Cleavage by Divergent CRISPR-Cas9 Enzymes
Joseph Bondy-Denomy, Alan R. Davidson  Trends in Microbiology 
The Mechanism of E. coli RNA Polymerase Regulation by ppGpp Is Suggested by the Structure of their Complex  Yuhong Zuo, Yeming Wang, Thomas A. Steitz 
Guide RNA Functional Modules Direct Cas9 Activity and Orthogonality
Volume 16, Issue 4, Pages (November 2004)
Volume 164, Issue 1, Pages (January 2016)
Dipali G. Sashital, Blake Wiedenheft, Jennifer A. Doudna 
RNAi: Prokaryotes Get in on the Act
Volume 25, Issue 6, Pages (March 2007)
Crystal Structures of RNase H Bound to an RNA/DNA Hybrid: Substrate Specificity and Metal-Dependent Catalysis  Marcin Nowotny, Sergei A. Gaidamakov, Robert.
Crystal Structure of a Y-Family DNA Polymerase in Action
Volume 28, Issue 6, Pages (December 2007)
RAS Proteins and Their Regulators in Human Disease
ACF Takes the Driver’s Seat
Volume 69, Issue 1, Pages e3 (January 2018)
Volume 30, Issue 3, Pages (May 2008)
Melissa S Jurica, Raymond J Monnat, Barry L Stoddard  Molecular Cell 
Structural Variation of Type I-F CRISPR RNA Guided DNA Surveillance
Volume 14, Issue 8, Pages (August 2006)
A CRISPR Approach to Gene Targeting
Mark Del Campo, Alan M. Lambowitz  Molecular Cell 
Regulatory RNAs in Bacteria
Nadja Heidrich, Jörg Vogel  Molecular Cell 
Box H/ACA Small Ribonucleoproteins
Volume 111, Issue 6, Pages (December 2002)
Structure of an RNA Silencing Complex of the CRISPR-Cas Immune System
Hayun Lee, Yi Zhou, David W. Taylor, Dipali G. Sashital  Molecular Cell 
Volume 23, Issue 4, Pages (April 2015)
Volume 29, Issue 6, Pages (March 2008)
Volume 52, Issue 3, Pages (November 2013)
Volume 34, Issue 3, Pages (May 2009)
Polymerases and the Replisome: Machines within Machines
Crystal Structures of RNase H Bound to an RNA/DNA Hybrid: Substrate Specificity and Metal-Dependent Catalysis  Marcin Nowotny, Sergei A. Gaidamakov, Robert.
Molecular Structures of Transcribing RNA Polymerase I
Structural Basis of 3′ End RNA Recognition and Exoribonucleolytic Cleavage by an Exosome RNase PH Core  Esben Lorentzen, Elena Conti  Molecular Cell 
The Biology of CRISPR-Cas: Backward and Forward
The Structure of T. aquaticus DNA Polymerase III Is Distinct from Eukaryotic Replicative DNA Polymerases  Scott Bailey, Richard A. Wing, Thomas A. Steitz 
Volume 65, Issue 6, Pages (March 2017)
DNA Targeting by a Minimal CRISPR RNA-Guided Cascade
Presentation transcript:

A Conserved Structural Chassis for Mounting Versatile CRISPR RNA-Guided Immune Responses  Ryan N. Jackson, Blake Wiedenheft  Molecular Cell  Volume 58, Issue 5, Pages 722-728 (June 2015) DOI: 10.1016/j.molcel.2015.05.023 Copyright © 2015 Elsevier Inc. Terms and Conditions

Figure 1 Re-rooting the CRISPR-Cas Phylogenetic Tree (A) Phylogenetic studies have identified three main CRISPR system types (I, II, and III) that are defined by the content and organization of cas genes and CRISPR loci. Type I and type III systems rely on large multi-subunit complexes composed of a CRISPR-derived RNA (crRNA) and several CRISPR-associated (cas) genes, whereas type II systems rely on two RNAs (i.e., tracrRNA and crRNA) and a single Cas9 protein. (B) Structures reveal a striking architectural similarity between the type I and type III surveillance complexes (PDB: 4QYZ, 3X1L), suggesting that these systems evolved from a common ancestor, while the Cas9 protein is structurally and evolutionarily distinct (PDB: 4UN3). (C) Type I complexes bind dsDNA and recruit transacting nuclease Cas3, while type III complexes bind single-stranded RNA with a 100-fold greater affinity than single-stranded DNA. Type III complexes cleave the RNA target using nuclease active sites that are positioned at discrete intervals along the backbone. These systems also cleave the non-template stand of transcriptionally active DNA. DNA cleavage relies on amino acids in the Cmr2 tail (purple) that are conserved in Cas10 family proteins. Type II systems rely on a single Cas9 protein that binds and cleaves dsDNA targets. Molecular Cell 2015 58, 722-728DOI: (10.1016/j.molcel.2015.05.023) Copyright © 2015 Elsevier Inc. Terms and Conditions

Figure 2 The Helical Backbone Is a Versatile Architectural Platform with Mechanistic Plasticity (A) The Cas7 protein from E. coli (type I-E) is shaped like a right hand. The palm of Cas7 binds the crRNA through non-sequence-specific interactions, and the thumb pierces the crRNA/DNA duplex (PDB: 4QYZ). (B) The Cascade backbone is composed of six interwoven Cas7 proteins (Cas7.1–7.6). The Cas7 thumbs distort the backbone geometry of the single-stranded DNA target, introducing ∼107° kinks at six-nucleotide intervals. (C) The Cmr4 protein from A. fulgidus is a Cas7 family protein with a “right hand” morphology. The crRNA is non-specifically bound by the palm, and the thumbs pierce the crRNA/target duplex at six-nucleotide intervals (PDB: 3X1L). (D) The catalytic residue (D31) on Cmr4 is adjacent to ∼56° kinks in the phosphate backbone. Conserved residues on Cmr5 (K144) and Cmr2 (K789) interact with the kinked out phosphate, and the side chain of a non-conserved residue (e.g., Cmr5E40) nudges the kinked bases toward the major groove of the adjacent duplex. Molecular Cell 2015 58, 722-728DOI: (10.1016/j.molcel.2015.05.023) Copyright © 2015 Elsevier Inc. Terms and Conditions

Figure 3 Tail Subunits Participate in the Destruction of DNA Targets (A) Type I and type III surveillance complexes share a multi-subunit, seahorse-shaped architecture. The tail subunits of both types are involved in DNA cleavage. The Cse1 protein of type I complexes recruits the trans-acting nuclease Cas3 for DNA cleavage, and the Csm2 protein of type III complexes is critical for DNA target degradation. (B) Structural comparison of the distinct tail subunits (PDB: 4TVX, 4W8Y). (C) Close-up of the conserved the GGDD motif and a coordinated manganese ion (green). (D) Close-up of the histidine-aspartate (HD) motif and two manganese ions (green) in the N-terminal domain. Molecular Cell 2015 58, 722-728DOI: (10.1016/j.molcel.2015.05.023) Copyright © 2015 Elsevier Inc. Terms and Conditions