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CryoEM structural determinations of Bacterial Restriction-Modification systems


EMSL Project ID
51306

Abstract

Nuclease enzymes are of enormous importance for a wide range of biological and biotechnology disciplines that require site-specific DNA manipulations. Bacterial R-M systems consist of a restriction endonuclease that specifically cleaves an unmodified nucleotide sequence, and a cognate DNA methyltransferase that protects the host DNA from cleavage. Such enzyme functions provide excellent models for studying specific protein-DNA interactions. Although most of well-understood restriction endonucleases correspond to relatively simple, small enzymes, the majority of such enzymes are instead comprised of large multi-subunit assemblages that coordinate the activity of multiple functional domains: DNA recognition, DNA cleavage, DNA methylation and often DNA translocation. To date, no structures of such large complexes have been determined. We are working towards Cryo-EM structural determinations of two such enzymes: BsaXI which is a large R-M complex that contains three functional subunits in a multi-subunit assemblage and DdrV, which contains the same functions encoded on a single polypeptide

Project Details

Start Date
2020-03-15
End Date
2021-03-17
Status
Closed

Team

Principal Investigator

Betty Shen
Institution
Fred Hutchinson Cancer Research Center

Co-Investigator(s)

Barry Stoddard
Institution
Fred Hutchinson Cancer Research Center

Team Members

Rose Marie Haynes
Institution
Environmental Molecular Sciences Laboratory

Madison Kennedy
Institution
Fred Hutchinson Cancer Research Center

Lindsey Doyle
Institution
Fred Hutchinson Cancer Research Center

Irina El Khoury
Institution
Environmental Molecular Sciences Laboratory