Structural basis for target site selection in RNA-guided DNA transposition systems.

Tytuł:: Structural basis for target site selection in RNA-guided DNA transposition systems.
Autorzy:: Park JU; Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA.
Tsai AW; Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA.
Mehrotra E; Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA.
Petassi MT; Department of Microbiology, Cornell University, Ithaca, NY 14853, USA.
Hsieh SC; Department of Microbiology, Cornell University, Ithaca, NY 14853, USA.
Ke A; Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA.
Peters JE; Department of Microbiology, Cornell University, Ithaca, NY 14853, USA. .
Kellogg EH; Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA. .
Źródło:: Science (New York, N.Y.) [Science] 2021 Aug 13; Vol. 373 (6556), pp. 768-774. Date of Electronic Publication: 2021 Jul 15.
Typ publikacji:: Journal Article; Research Support, N.I.H., Extramural
Język:: English
Imprint Name(s):: Publication: : Washington, DC : American Association for the Advancement of Science
Original Publication: New York, N.Y. : [s.n.] 1880-
MeSH Terms:: DNA Transposable Elements*
Bacterial Proteins/*chemistry
CRISPR-Associated Proteins/*chemistry
Cyanobacteria/*chemistry
Adenosine Diphosphate/metabolism ; Adenosine Triphosphate/metabolism ; Bacterial Proteins/metabolism ; CRISPR-Associated Proteins/metabolism ; Cryoelectron Microscopy ; Cyanobacteria/genetics ; Cyanobacteria/metabolism ; DNA, Bacterial/metabolism ; Models, Molecular ; Protein Conformation ; Protein Folding ; RNA, Bacterial/metabolism ; Transposases/chemistry ; Transposases/metabolism
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Grant Information:: R01 GM144566 United States GM NIGMS NIH HHS; R01 GM129118 United States GM NIGMS NIH HHS; R00 GM124463 United States GM NIGMS NIH HHS; R21 AI148941 United States AI NIAID NIH HHS; R35 GM118174 United States GM NIGMS NIH HHS
Substance Nomenclature:: 0 (Bacterial Proteins)
0 (CRISPR-Associated Proteins)
0 (DNA Transposable Elements)
0 (DNA, Bacterial)
0 (RNA, Bacterial)
61D2G4IYVH (Adenosine Diphosphate)
8L70Q75FXE (Adenosine Triphosphate)
EC 2.7.7.- (Transposases)
SCR Organism:: Scytonema hofmannii
Entry Date(s):: Date Created: 20210813 Date Completed: 20210820 Latest Revision: 20220716
Update Code:: 20240105
PubMed Central ID:: PMC9080059
DOI:: 10.1126/science.abi8976
PMID:: 34385391
: Czasopismo naukowe

Pełny tekst

CRISPR-associated transposition systems allow guide RNA-directed integration of a single DNA cargo in one orientation at a fixed distance from a programmable target sequence. We used cryo-electron microscopy (cryo-EM) to define the mechanism that underlies this process by characterizing the transposition regulator, TnsC, from a type V-K CRISPR-transposase system. In this scenario, polymerization of adenosine triphosphate-bound TnsC helical filaments could explain how polarity information is passed to the transposase. TniQ caps the TnsC filament, representing a universal mechanism for target information transfer in Tn7/Tn7-like elements. Transposase-driven disassembly establishes delivery of the element only to unused protospacers. Finally, TnsC transitions to define the fixed point of insertion, as revealed by structures with the transition state mimic ADP•AlF 3 These mechanistic findings provide the underpinnings for engineering CRISPR-associated transposition systems for research and therapeutic applications.
(Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

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