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10.1016/j.cell.2017.06.012 http://scihub22266oqcxt.onion/10.1016/j.cell.2017.06.012 C5841471!5841471!28666122     free     free     free Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 253.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Cell 2017 ; 170 (1): 48-60.e11 Nephropedia Template TP {{tp|p=28666122|t=2017. Structure basis for directional R-loop formation and substrate handover mechanisms in Type I CRISPR-Cas system |pdf=|usr=}}{{28666122}} gab.com Text Structure basis for directional R-loop formation and substrate handover mechanisms in Type I CRISPR-Cas system JO: Cell http://www.kidney.de/pget.php?&tw=1&pmid=28666122 #FOAvip Type I CRISPR systems feature a sequential dsDNA target searching and degradation process, by crRNA-displaying Cascade and nuclease-helicase fusion enzyme Cas3, respectively. Here we present two cryo-EM snapshots of the Thermobifida fusca Type I-E Cascade: 1) unwinding 11-bp of dsDNA at the seed-sequence region to scout for sequence complementarity, and 2) further unwinding of the entire protospacer to form a full R-loop. These structures provide the much-needed temporal and spatial resolution to resolve key mechanistic steps leading to Cas3 recruitment. In the early steps, PAM recognition causes severe DNA bending, leading to spontaneous DNA unwinding to form a seed-bubble. The full R-loop formation triggers conformational changes in Cascade, licensing Cas3 to bind. The same process also generates a bulge in the non-target DNA strand, enabling its handover to Cas3 for cleavage. The combination of both negative and positive checkpoints ensures stringent yet efficient target degradation in Type I CRISPR-Cas systems. Twit Text FOAVip Structure basis for directional R-loop formation and substrate handover mechanisms in Type I CRISPR-Cas system ????Cell http://www.kidney.de/pget.php?&tw=1&pmid=28666122 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=28666122 #FOAvip English Wikipedia <ref>{{Cite pmid|28666122}}</ref> Structure basis for directional R-loop formation and substrate handover mechanisms in Type I CRISPR-Cas system #MMPMID28666122 Xiao Y; Luo M; Hayes RP; Kim J; Ng S; Ding F; Liao M; Ke A Cell 2017[Jun]; 170 (1): 48-60.e11 PMID28666122show ga Type I CRISPR systems feature a sequential dsDNA target searching and degradation process, by crRNA-displaying Cascade and nuclease-helicase fusion enzyme Cas3, respectively. Here we present two cryo-EM snapshots of the Thermobifida fusca Type I-E Cascade: 1) unwinding 11-bp of dsDNA at the seed-sequence region to scout for sequence complementarity, and 2) further unwinding of the entire protospacer to form a full R-loop. These structures provide the much-needed temporal and spatial resolution to resolve key mechanistic steps leading to Cas3 recruitment. In the early steps, PAM recognition causes severe DNA bending, leading to spontaneous DNA unwinding to form a seed-bubble. The full R-loop formation triggers conformational changes in Cascade, licensing Cas3 to bind. The same process also generates a bulge in the non-target DNA strand, enabling its handover to Cas3 for cleavage. The combination of both negative and positive checkpoints ensures stringent yet efficient target degradation in Type I CRISPR-Cas systems. äStructure basis for directional R-loop formation and substrate handove(epmc).pdf DeepDyve Pubget Overpricing