This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Heimer, S. R.
Right arrow Articles by Mobley, H. L. T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Heimer, S. R.
Right arrow Articles by Mobley, H. L. T.

 Previous Article  |  Next Article 

Journal of Bacteriology, February 2001, p. 1423-1433, Vol. 183, No. 4
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.4.1423-1433.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Interaction of Proteus mirabilis Urease Apoenzyme and Accessory Proteins Identified with Yeast Two-Hybrid Technology

Susan R. Heimer and Harry L. T. Mobley*

Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, Maryland 21201

Received 1 September 2000/Accepted 16 November 2000

Proteus mirabilis, a gram-negative bacterium associated with complicated urinary tract infections, produces a metalloenzyme urease which hydrolyzes urea to ammonia and carbon dioxide. The apourease is comprised of three structural subunits, UreA, UreB, and UreC, assembled as a homotrimer of individual UreABC heterotrimers (UreABC)3. To become catalytically active, apourease acquires divalent nickel ions through a poorly understood process involving four accessory proteins, UreD, UreE, UreF, and UreG. While homologues of UreD, UreF, and UreG have been copurified with apourease, it remains unclear specifically how these polypeptides associate with the apourease or each other. To identify interactions among P. mirabilis accessory proteins, in vitro immunoprecipitation and in vivo yeast two-hybrid assays were employed. A complex containing accessory protein UreD and structural protein UreC was isolated by immunoprecipitation and characterized with immunoblots. This association occurs independently of coaccessory proteins UreE, UreF, and UreG and structural protein UreA. In a yeast two-hybrid screen, UreD was found to directly interact in vivo with coaccessory protein UreF. Unique homomultimeric interactions of UreD and UreF were also detected in vivo. To substantiate the study of urease proteins with a yeast two-hybrid assay, previously described UreE dimers and homomultimeric UreA interactions among apourease trimers were confirmed in vivo. Similarly, a known structural interaction involving UreA and UreC was also verified. This report suggests that in vivo, P. mirabilis UreD may be important for recruitment of UreF to the apourease and that crucial homomultimeric associations occur among these accessory proteins.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD 21201. Phone: (410) 706-0466. Fax: (410) 706-6751. E-mail: hmobley{at}umaryland.edu.

Journal of Bacteriology, February 2001, p. 1423-1433, Vol. 183, No. 4
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.4.1423-1433.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Booth, N. J., Beekman, J. B., Thune, R. L. (2009). Edwardsiella ictaluri Encodes an Acid-Activated Urease That Is Required for Intracellular Replication in Channel Catfish (Ictalurus punctatus) Macrophages. Appl. Environ. Microbiol. 75: 6712-6720 [Abstract] [Full Text]  
  • Kim, J. K., Mulrooney, S. B., Hausinger, R. P. (2006). The UreEF Fusion Protein Provides a Soluble and Functional Form of the UreF Urease Accessory Protein. J. Bacteriol. 188: 8413-8420 [Abstract] [Full Text]  
  • Zambelli, B., Stola, M., Musiani, F., De Vriendt, K., Samyn, B., Devreese, B., Van Beeumen, J., Turano, P., Dikiy, A., Bryant, D. A., Ciurli, S. (2005). UreG, a Chaperone in the Urease Assembly Process, Is an Intrinsically Unstructured GTPase That Specifically Binds Zn2+. J. Biol. Chem. 280: 4684-4695 [Abstract] [Full Text]  
  • Burall, L. S., Harro, J. M., Li, X., Lockatell, C.V., Himpsl, S. D., Hebel, J. R., Johnson, D. E., Mobley, H. L. T. (2004). Proteus mirabilis Genes That Contribute to Pathogenesis of Urinary Tract Infection: Identification of 25 Signature-Tagged Mutants Attenuated at Least 100-Fold. Infect. Immun. 72: 2922-2938 [Abstract] [Full Text]  
  • Chang, Z., Kuchar, J., Hausinger, R. P. (2004). Chemical Cross-linking and Mass Spectrometric Identification of Sites of Interaction for UreD, UreF, and Urease. J. Biol. Chem. 279: 15305-15313 [Abstract] [Full Text]  
  • Mehta, N., Olson, J. W., Maier, R. J. (2003). Characterization of Helicobacter pylori Nickel Metabolism Accessory Proteins Needed for Maturation of both Urease and Hydrogenase. J. Bacteriol. 185: 726-734 [Abstract] [Full Text]  
  • Dattelbaum, J. D., Lockatell, C. V., Johnson, D. E., Mobley, H. L. T. (2003). UreR, the Transcriptional Activator of the Proteus mirabilis Urease Gene Cluster, Is Required for Urease Activity and Virulence in Experimental Urinary Tract Infections. Infect. Immun. 71: 1026-1030 [Abstract] [Full Text]  
  • Voland, P., Weeks, D. L., Marcus, E. A., Prinz, C., Sachs, G., Scott, D. (2003). Interactions among the seven Helicobacter pylori proteins encoded by the urease gene cluster. Am. J. Physiol. Gastrointest. Liver Physiol. 284: G96-G106 [Abstract] [Full Text]  
  • Heimer, S. R., Welch, R. A., Perna, N. T., Posfai, G., Evans, P. S., Kaper, J. B., Blattner, F. R., Mobley, H. L. T. (2002). Urease of Enterohemorrhagic Escherichia coli: Evidence for Regulation by Fur and a trans-Acting Factor. Infect. Immun. 70: 1027-1031 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»