Structure-function studies of Escherichia coli RnlA reveal a novel toxin structure involved in bacteriophage resistance.

Abstract:

:Escherichia coli RnlA-RnlB is a newly identified toxin-antitoxin (TA) system that plays a role in bacteriophage resistance. RnlA functions as a toxin with mRNA endoribonuclease activity and the cognate antitoxin RnlB inhibits RnlA toxicity in E. coli cells. Interestingly, T4 phage encodes the antitoxin Dmd, which acts against RnlA to promote its own propagation, suggesting that RnlA-Dmd represents a novel TA system. Here, we have determined the crystal structure of RnlA refined to 2.10  (Dmd-binding domain), which is an organization not previously observed among known toxin structures. Small-angle X-ray scattering (SAXS) analysis revealed that RnlA forms a dimer in solution via interactions between the DBDs from both monomers. The in vitro and in vivo functional studies showed that among the three domains, only the DBD is responsible for recognition and inhibition by Dmd and subcellular location of RnlA. In particular, the helix located at the C-terminus of DBD plays a vital role in binding Dmd. Our comprehensive studies reveal the key region responsible for RnlA toxicity and provide novel insights into its structure-function relationship.

journal_name

Mol Microbiol

journal_title

Molecular microbiology

authors

Wei Y,Gao ZQ,Otsuka Y,Naka K,Yonesaki T,Zhang H,Dong YH

doi

10.1111/mmi.12409

subject

Has Abstract

pub_date

2013-12-01 00:00:00

pages

956-65

issue

5

eissn

0950-382X

issn

1365-2958

journal_volume

90

pub_type

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