The Structure of Xis Reveals the Basis for Filament Formation and Insight into DNA Bending Within a Mycobacteriophage Intasome

Overview

Journal J Mol Biol

Publisher Elsevier

Specialties Microbiology
Molecular Biology

Date 2013 Oct 12

PMID 24112940

Citations 9

Authors

Shweta Singh

Joseph G Plaks

Nicholas J Homa

Christopher G Amrich

Annie Heroux

Graham F Hatfull

Andrew P VanDemark

Affiliations

Soon will be listed here.

Abstract

The recombination directionality factor, Xis, is a DNA bending protein that determines the outcome of integrase-mediated site-specific recombination by redesign of higher-order protein-DNA architectures. Although the attachment site DNA of mycobacteriophage Pukovnik is likely to contain four sites for Xis binding, Xis crystals contain five subunits in the asymmetric unit, four of which align into a Xis filament and a fifth that is generated by an unusual domain swap. Extensive intersubunit contacts stabilize a bent filament-like arrangement with Xis monomers aligned head to tail. The structure implies a DNA bend of ~120°, which is in agreement with DNA bending measured in vitro. Formation of attR-containing intasomes requires only Int and Xis, distinguishing Pukovnik from lambda. Therefore, we conclude that, in Pukovnik, Xis-induced DNA bending is sufficient to promote intramolecular Int-mediated bridges during intasome formation.

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