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Journal of Bacteriology, February 2002, p. 629-635, Vol. 184, No. 3
0021-9193/01/$04.00+0     DOI: 10.1128/JB.184.3.629-635.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Evidence for Direct Protein-Protein Interaction between Members of the Enterobacterial Hha/YmoA and H-NS Families of Proteins

J. M. Nieto, C. Madrid, E. Miquelay, J. L. Parra, S. Rodríguez, and A. Juárez*

Departament de Microbiologia, Facultat de Biologia, Universitat de Barcelona, 08028 Barcelona, Spain

Received 10 August 2001/ Accepted 7 November 2001

Escherichia coli nucleoid-associated H-NS protein interacts with the Hha protein, a member of a new family of global modulators that also includes the YmoA protein from Yersinia enterocolitica. This interaction has been found to be involved in the regulation of the expression of the toxin {alpha}-hemolysin. In this study, we further characterize the interaction between H-NS and Hha. We show that the presence of DNA in preparations of copurified His-Hha and H-NS is not directly implicated in the interaction between the proteins. The precise molecular mass of the H-NS protein retained by Hha, obtained by mass spectrometry analysis, does not show any posttranslational modification other than removal of the N-terminal Met residue. We constructed an H-NS-His recombinant protein and found that, as expected, it interacts with Hha. We used a Ni2+-nitrilotriacetic acid agarose method for affinity chromatography copurification of proteins to identify the H-NS protein of Y. enterocolitica. We constructed a six-His-YmoA recombinant protein derived from YmoA, the homologue of Hha in Y. enterocolitica, and found that it interacts with Y. enterocolitica H-NS. We also cloned and sequenced the hns gene of this microorganism. In the course of these experiments we found that His-YmoA can also retain H-NS from E. coli. We also found that the hns gene of Y. enterocolitica can complement an hns mutation of E. coli. Finally, we describe for the first time systematic characterization of missense mutant alleles of hha and truncated Hha' proteins, and we report a striking and previously unnoticed similarity of the Hha family of proteins to the oligomerization domain of the H-NS proteins.

* Corresponding author. Mailing address: Departament de Microbiologia, Facultat de Biologia, Universitat de Barcelona, Avda. Diagonal, 645, 08028 Barcelona, Spain. Phone: 3493 4034624. Fax: 3493 4034629. E-mail: juarez{at}porthos.bio.ub.es.

Journal of Bacteriology, February 2002, p. 629-635, Vol. 184, No. 3
0021-9193/01/$04.00+0     DOI: 10.1128/JB.184.3.629-635.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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