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Journal of Bacteriology, May 2005, p. 3151-3157, Vol. 187, No. 9
0021-9193/05/$08.00+0     doi:10.1128/JB.187.9.3151-3157.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

RNase/Anti-RNase Activities of the Bacterial parD Toxin-Antitoxin System

Ana J. Muñoz-Gómez,1,{dagger} Marc Lemonnier,1,{dagger}* Sandra Santos-Sierra,1,{ddagger} Alfredo Berzal-Herranz,2 and Ramón Díaz-Orejas1

Department of Molecular Microbiology, Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, E-28040 Madrid, Spain,1 Instituto de Parasitología y Biomedicina "López-Neyra", CSIC, Parque Tecnológico de Ciencias de la Salud, Avda. del Conocimiento s/n, Armilla, E-18100 Granada, Spain2

Received 20 December 2004/ Accepted 25 January 2005

The bacterial parD toxin-antitoxin system of plasmid R1 encodes two proteins, the Kid toxin and its cognate antitoxin, Kis. Kid cleaves RNA and inhibits protein synthesis and cell growth in Escherichia coli. Here, we show that Kid promotes RNA degradation and inhibition of protein synthesis in rabbit reticulocyte lysates. These new activities of the Kid toxin were counteracted by the Kis antitoxin and were not displayed by the KidR85W variant, which is nontoxic in E. coli. Moreover, while Kid cleaved single- and double-stranded RNA with a preference for UAA or UAC triplets, KidR85W maintained this sequence preference but hardly cleaved double-stranded RNA. Kid was formerly shown to inhibit DNA replication of the ColE1 plasmid. Here we provide in vitro evidence that Kid cleaves the ColE1 RNA II primer, which is required for the initiation of ColE1 replication. In contrast, KidR85W did not affect the stability of RNA II, nor did it inhibit the in vitro replication of ColE1. Thus, the endoribonuclease and the cytotoxic and DNA replication-inhibitory activities of Kid seem tightly correlated. We propose that the spectrum of action of this toxin extends beyond the sole inhibition of protein synthesis to control a broad range of RNA-regulated cellular processes.

* Corresponding author. Mailing address: Department of Molecular Microbiology, Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, E-28040 Madrid, Spain. Phone: +34 91 837 31 12. Fax: +34 91 536 04 32. E-mail: mlm{at}cib.csic.es.

{dagger} These authors contributed equally to this work.

{ddagger} Present address: Institut für Biochemische Pharmakologie, Universität Innsbruck, Peter Mayr Strasse 1, A-6020 Innsbruck, Austria.

Journal of Bacteriology, May 2005, p. 3151-3157, Vol. 187, No. 9
0021-9193/05/$08.00+0     doi:10.1128/JB.187.9.3151-3157.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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