Organization of the BcgI Restriction-modification Protein for the Cleavage of Eight Phosphodiester Bonds in DNA

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2012 Nov 14

PMID 23147005

Citations 9

Authors

Rachel M Smith

Jacqueline J T Marshall

Alistair J Jacklin

Susan E Retter

Stephen E Halford

Frank Sobott

Affiliations

Soon will be listed here.

Abstract

Type IIB restriction-modification systems, such as BcgI, feature a single protein with both endonuclease and methyltransferase activities. Type IIB nucleases require two recognition sites and cut both strands on both sides of their unmodified sites. BcgI cuts all eight target phosphodiester bonds before dissociation. The BcgI protein contains A and B polypeptides in a 2:1 ratio: A has one catalytic centre for each activity; B recognizes the DNA. We show here that BcgI is organized as A(2)B protomers, with B at its centre, but that these protomers self-associate to assemblies containing several A(2)B units. Moreover, like the well known FokI nuclease, BcgI bound to its site has to recruit additional protomers before it can cut DNA. DNA-bound BcgI can alternatively be activated by excess A subunits, much like the activation of FokI by its catalytic domain. Eight A subunits, each with one centre for nuclease activity, are presumably needed to cut the eight bonds cleaved by BcgI. Its nuclease reaction may thus involve two A(2)B units, each bound to a recognition site, with two more A(2)B units bridging the complexes by protein-protein interactions between the nuclease domains.

Citing Articles

Structure, subunit organization and behavior of the asymmetric Type IIT restriction endonuclease BbvCI.

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A model for the evolution of prokaryotic DNA restriction-modification systems based upon the structural malleability of Type I restriction-modification enzymes.

Bower E, Cooper L, Roberts G, White J, Luyten Y, Morgan R Nucleic Acids Res. 2018; 46(17):9067-9080.

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Modification-dependent restriction endonuclease, MspJI, flips 5-methylcytosine out of the DNA helix.

Horton J, Wang H, Mabuchi M, Zhang X, Roberts R, Zheng Y Nucleic Acids Res. 2014; 42(19):12092-101.

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Structure of 5-hydroxymethylcytosine-specific restriction enzyme, AbaSI, in complex with DNA.

Horton J, Borgaro J, Griggs R, Quimby A, Guan S, Zhang X Nucleic Acids Res. 2014; 42(12):7947-59.

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