Abstract：
:Bacteria acquire memory of viral invaders by incorporating invasive DNA sequence elements into the host CRISPR locus, generating a new spacer within the CRISPR array. We report on the structures of Cas1-Cas2-dual-forked DNA complexes in an effort toward understanding how the protospacer is sampled prior to insertion into the CRISPR locus. Our study reveals a protospacer DNA comprising a 23-bp duplex bracketed by tyrosine residues, together with anchored flanking 3' overhang segments. The PAM-complementary sequence in the 3' overhang is recognized by the Cas1a catalytic subunits in a base-specific manner, and subsequent cleavage at positions 5 nt from the duplex boundary generates a 33-nt DNA intermediate that is incorporated into the CRISPR array via a cut-and-paste mechanism. Upon protospacer binding, Cas1-Cas2 undergoes a significant conformational change, generating a flat surface conducive to proper protospacer recognition. Here, our study provides important structure-based mechanistic insights into PAM-dependent spacer acquisition.
journal_name
Celljournal_title
Cellauthors
Wang J,Li J,Zhao H,Sheng G,Wang M,Yin M,Wang Ydoi
10.1016/j.cell.2015.10.008subject
Has Abstractpub_date
2015-11-05 00:00:00pages
840-53issue
4eissn
0092-8674issn
1097-4172pii
S0092-8674(15)01321-5journal_volume
163pub_type
杂志文章相关文献
CELL文献大全abstract：:The crystal structure of the β2-adrenergic receptor (β2AR) bound to the G protein adenylyl cyclase stimulatory G protein (Gs) captured the complex in a nucleotide-free state (β2AR-Gsempty). Unfortunately, the β2AR-Gsempty complex does not provide a clear explanation for G protein coupling specificity. Evidence from se...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/j.cell.2019.04.021
更新日期：2019-05-16 00:00:00
abstract：:Breast cancer (BC) comprises multiple distinct subtypes that differ genetically, pathologically, and clinically. Here, we describe a robust protocol for long-term culturing of human mammary epithelial organoids. Using this protocol, >100 primary and metastatic BC organoid lines were generated, broadly recapitulating t...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/j.cell.2017.11.010
更新日期：2018-01-11 00:00:00
abstract：:The protein Par-4 acts in the cytoplasm to trigger cell death signaling via caspase activation and the mitochondrial release of cytochrome c. Burikhanov et al. (2009) now provide surprising evidence that Par-4 can also promote apoptosis from outside the cell, after its secretion in response to endoplasmic reticulum st...
journal_title：Cell
pub_type： 评论,杂志文章
doi：10.1016/j.cell.2009.07.007
更新日期：2009-07-23 00:00:00
abstract：:Biomechanical stress is a major stimulus for cardiac hypertrophy and the transition to heart failure. By generating mice that harbor a ventricular restricted knockout of the gp130 cytokine receptor via Cre-IoxP-mediated recombination, we demonstrate a critical role for a gp130-dependent myocyte survival pathway in the...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/s0092-8674(00)80729-1
更新日期：1999-04-16 00:00:00
abstract：:The mammalian alpha- and beta--globin genes are thought to have evolved from a common ancestral sequence by a duplication event that occurred over 500 million years ago. We have now determined the entire nucleotide sequence of a cloned mouse alpha--globin gene, including regions that flank and interrupt the coding seq...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/0092-8674(79)90139-9
更新日期：1979-11-01 00:00:00
abstract：:We used clinical tissue from lethal metastatic castration-resistant prostate cancer (CRPC) patients obtained at rapid autopsy to evaluate diverse genomic, transcriptomic, and phosphoproteomic datasets for pathway analysis. Using Tied Diffusion through Interacting Events (TieDIE), we integrated differentially expressed...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/j.cell.2016.07.007
更新日期：2016-08-11 00:00:00
abstract：:Activation of the cyclin-dependent kinases to promote cell cycle progression requires their association with cyclins as well as phosphorylation of a threonine (residue 161 in human p34cdc2). This phosphorylation is carried out by CAK, the Cdk-activating kinase. We have purified and cloned CAK from S. cerevisiae. Unlik...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/s0092-8674(00)80129-4
更新日期：1996-08-23 00:00:00
abstract：:We have identified in mice an allele of a new T cell receptor V beta gene, V beta 17a, whose product is bound by the monoclonal antibody KJ23a. Over 90% of T cell hybridomas prepared from V beta 17a+ T cells of SWR mice respond to allogeneic forms of the IE class II MHC protein, indicating that V beta 17a has an appre...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/0092-8674(87)90567-8
更新日期：1987-04-24 00:00:00
abstract：:We propose a mechanism for the priming of influenza viral RNA transcription by capped RNAs in which specific 5'-terminal fragments are cleaved from the capped RNAs by a virion-associated endonuclease. These fragments would serve as the actual primers for the initiation of transcription by the initial incorporation by ...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/0092-8674(81)90449-9
更新日期：1981-03-01 00:00:00
abstract：:In many organisms, meiotic chromosomes are bundled at their telomeres to form a "bouquet" arrangement. The bouquet formation plays an important role in homologous chromosome pairing and therefore progression of meiosis. As meiotic telomere clustering occurs in response to mating pheromone signaling in fission yeast, w...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/j.cell.2006.01.048
更新日期：2006-04-07 00:00:00
abstract：:The MYC oncoproteins are thought to stimulate tumor cell growth and proliferation through amplification of gene transcription, a mechanism that has thwarted most efforts to inhibit MYC function as potential cancer therapy. Using a covalent inhibitor of cyclin-dependent kinase 7 (CDK7) to disrupt the transcription of a...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/j.cell.2014.10.024
更新日期：2014-11-20 00:00:00
abstract：:Symbiotic bacteria are important in animal hosts, but have been largely overlooked as they have proved difficult to culture in the laboratory. Approaches such as comparative genomics and real-time PCR have provided insights into the molecular mechanisms that underpin symbiont-host interactions. Studies on the heritabl...
journal_title：Cell
pub_type： 杂志文章,评审
doi：10.1016/j.cell.2006.07.014
更新日期：2006-08-11 00:00:00
abstract：:The transcriptional induction of heat shock genes is mediated by the heat shock transcription factor (HSF). We have investigated the oligomerization state of HSF from S. cerevisiae and find that it forms a trimer in solution and when bound to DNA. Trimerization is mediated by a region of HSF that, like the leucine zip...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/0092-8674(89)90604-1
更新日期：1989-12-01 00:00:00
abstract：:We have investigated RNA recombination among poliovirus genomes by analyzing both intratypic and intertypic recombinant crosses involving the same defined genetic markers. Sequence analysis of the recombinant junctions of 13 nonsibling intertypic recombinants showed that intertypic RNA recombination is not site-specif...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/0092-8674(86)90600-8
更新日期：1986-11-07 00:00:00
abstract：:Kinetochores connect centromeric nucleosomes with mitotic-spindle microtubules through conserved, cross-interacting protein subassemblies. In budding yeast, the heterotetrameric MIND complex (Mtw1, Nnf1, Nsl1, Dsn1), ortholog of the metazoan Mis12 complex, joins the centromere-proximal components, Mif2 and COMA, with ...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/j.cell.2016.10.011
更新日期：2016-11-03 00:00:00
abstract：:The pathway of RNA polymerase entry at the lac promoter was studied by investigating the relationship between the promoter and a weak, overlapping polymerase interaction site (P2). If polymerase is made to enter the DNA by binding in vitro at this P2 site, cyclic AMP receptor protein (CRP) actively removes polymerase ...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/0092-8674(85)90250-8
更新日期：1985-12-01 00:00:00
abstract：:Multiple proteins act co-operatively in mammalian clathrin-mediated endocytosis (CME) to generate endocytic vesicles from the plasma membrane. The principles controlling the activation and organization of the actin cytoskeleton during mammalian CME are, however, not fully understood. Here, we show that the protein FCH...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/j.cell.2018.05.020
更新日期：2018-07-12 00:00:00
abstract：:Spinocerebellar ataxia type 1 (SCA1) is one of nine inherited, typically adult onset, polyglutamine neurodegenerative diseases. To examine whether development impacts SCA1, we used a conditional transgenic mouse model of SCA1 to delay the postnatal expression of mutant ATXN1 until after completion of cerebellar develo...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/j.cell.2006.09.036
更新日期：2006-11-17 00:00:00
abstract：:During asymmetric cell divisions, mitotic spindles align along the axis of polarization. In invertebrates, spindle positioning requires Pins or related proteins and a G protein alpha subunit. A mammalian Pins, called LGN, binds Galphai and also interacts through an N-terminal domain with the microtubule binding protei...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/j.cell.2004.10.028
更新日期：2004-11-12 00:00:00
abstract：:During development, the precise relationships between transcription and chromatin modifications often remain unclear. We use the X chromosome inactivation (XCI) paradigm to explore the implication of chromatin changes in gene silencing. Using female mouse embryonic stem cells, we initiate XCI by inducing Xist and then...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/j.cell.2018.11.041
更新日期：2019-01-10 00:00:00
abstract：:The crystal structure of the snake toxin fasciculin, bound to mouse acetylcholinesterase (mAChE), at 3.2 A resolution reveals a synergistic three-point anchorage consistent with the picomolar dissociation constant of the complex. Loop II of fasciculin contains a cluster of hydrophobic residues that interact with the p...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/0092-8674(95)90128-0
更新日期：1995-11-03 00:00:00
abstract：:Actomyosin retrograde flow underlies the contraction essential for cell motility. Retrograde flow in both lamellipodia and lamella is required for membrane protrusion and for force generation by coupling to cell adhesion. We report that the Rac/Cdc42-binding kinase MRCK and myosin II-related MYO18A linked by the adapt...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/j.cell.2008.09.018
更新日期：2008-10-03 00:00:00
abstract：:Although a great deal is known biochemically about peroxiredoxins (Prxs), little is known about their real physiological function. We show here that two cytosolic yeast Prxs, cPrxI and II, which display diversity in structure and apparent molecular weights (MW), can act alternatively as peroxidases and molecular chape...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/j.cell.2004.05.002
更新日期：2004-05-28 00:00:00
abstract：:Hox homeodomain proteins are developmental regulators that determine body plan in a variety of organisms. A majority of the vertebrate Hox proteins bind DNA as heterodimers with the Pbx1 homeodomain protein. We report here the 2.35 A structure of a ternary complex containing a human HoxB1-Pbx1 heterodimer bound to DNA...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/s0092-8674(00)80662-5
更新日期：1999-02-19 00:00:00
abstract：:We have compared the chromatin structure in the active and inactive states at loci encoding the major heat shock protein in Drosophila. DNAase I and micrococcal nuclease were used as probes of higher order organization and nucleosomal integrity. Such integrity is gauged here by the characteristic pattern of discrete D...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/0092-8674(79)90096-5
更新日期：1979-04-01 00:00:00
abstract：:In ciliates, small RNAs have been shown to target foreign sequences for silencing via elimination from the somatic genome. Fang et al. now reveal a set of Piwi-interacting RNAs (piRNAs) in Oxytricha trifallax that likewise enable genomic self versus nonself discrimination, this time by specifying self sequences for ge...
journal_title：Cell
pub_type： 评论,杂志文章
doi：10.1016/j.cell.2012.11.032
更新日期：2012-12-07 00:00:00
abstract：:The LIM domain protein rbtn2 is associated with T cell acute leukemias. We demonstrate that rbtn2 is a nuclear protein expressed in the erythroid lineage in vivo, and using homologous recombination, we show that it is essential for erythroid development in mice. The homozygous rbtn2 null mutation leads to failure of y...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/0092-8674(94)90571-1
更新日期：1994-07-15 00:00:00
abstract：:The HIV auxiliary protein Vpr potently blocks the cell cycle at the G2/M transition. Here, we show that G2/M arrest results from untimely activation of the structure-specific endonuclease (SSE) regulator SLX4 complex (SLX4com) by Vpr, a process that requires VPRBP-DDB1-CUL4 E3-ligase complex. Direct interaction of Vpr...
journal_title：Cell
pub_type： 杂志文章
doi：10.1016/j.cell.2013.12.011
更新日期：2014-01-16 00:00:00
abstract：:Smad4 is an important mediator of signaling by TGFbeta family members that is presumed to be controlled by the phosphorylation status of its partners, the receptor-regulated R-Smads. In this issue, Dupont et al. (2009) now reveal that Smad4 itself is subject to cycles of ubiquitination and deubiquitination that regula...
journal_title：Cell
pub_type： 评论,杂志文章
doi：10.1016/j.cell.2008.12.028
更新日期：2009-01-09 00:00:00
abstract：:During metastasis, migrating breast cancer stem cells undergo a loss of polarity leading to an epithelial-to-mesenchymal transition (EMT). Gupta et al. (2009) use this attribute of cancer stem cells to develop a high-throughput screen, which successfully identifies small molecules that specifically inhibit cancer stem...
journal_title：Cell
pub_type： 评论,杂志文章
doi：10.1016/j.cell.2009.08.007
更新日期：2009-08-21 00:00:00
