Skip Navigation



Journal of Biochemistry Advance Access published online on April 3, 2008
Journal of Biochemistry, doi:10.1093/jb/mvn044
This Article
Right arrow Advance Access manuscript (PDF)
Right arrow Supplementary Data
Right arrow All Versions of this Article:
144/1/63    most recent
mvn044v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
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 Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Request Permissions
Google Scholar
Right arrow Articles by Rigden, M. D.
Right arrow Articles by Dillon, J.-A. R.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Rigden, M. D.
Right arrow Articles by Dillon, J.-A. R.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© 2008 The Japanese Biochemical Society

Identification of the Coiled-coil Domains of Enterococcus faecalis DivIVA that Mediate Oligomerization and Their Importance for Biological Function

Marc D. Rigden1, Cherise Baier2, Sandra Ramirez-Arcos1,§, Mingmin Liao2,3, Monica Wang3,4 and Jo-Anne R. Dillon1,2,3,4,¥

1Department of Biochemistry, Microbiology & Immunology, University of Ottawa, 451 Smyth Road, Ottawa, Ontario K1H 8M5, Canada
2Department of Microbiology & Immunology, University of Saskatchewan, 107 Wiggins Road, Saskatoon, Saskatchewan S7N 5E5
3Vaccine & Infectious Disease Organization, University of Saskatchewan, 120 Veterinary Road, Saskatoon, Saskatchewan S7N 5E3
4Department of Biology, University of Saskatchewan, 112 Science Place, Saskatoon, Saskatchewan, S7N 5E2, Canada

¥Current address and corresponding author: Prof. Jo-Anne R. Dillon. College of Arts and Science, University of Saskatchewan, Room 226, Arts Building 9 Campus Drive Saskatoon, Saskatchewan, Canada S7N 5A5. Tel: 1-306-966-4232; Fax: 1-306-966-8839, Email: j.dillon{at}usask.ca

Received October 24, 2007; Accepted March 13, 2008


  Abstract

Summary

Bacillus subtilis (Bs) DivIVA comprises coiled-coil structures and self-associates forming a 10-12 mer complex in vitro. Using bioinformatic approaches, we determined that Enterococcus faecalis (Ef) DivIVA comprises four coiled-coil domains, one at the N-terminus, the second and the third in the central region of the protein and the fourth at the C-terminus. We determined that DivIVAEf self-interacts and forms a 10-12 multimeric complex. Point mutations or deletions of the central regions predicted bioinformatically to disrupt the coiled-coil structures either eliminated or weakened DivIVAEf self-interaction and reduced oligomerization. Mutations disrupting the N- and C-terminal coiled-coils of DivIVAEf did not affect DivIVAEf oligomerization. The introduction of DivIVAEf mutations to both the N-terminal and the central coiled-coil domains were lethal unless rescued by expressing wild type DivIVAEf in trans. E. faecalis cells expressing these mutations displayed aberrant cell morphology, indicating disruption of the normal cell division phenotype. The results in E. faecalis also indicate that both the N-terminal and the central coiled-coil structures of DivIVAEf are indispensable for proper biological function. Overexpression of wild type DivIVAEf in both rod-shaped and round Escherichia coli cells resulted in morphological changes, while the overexpression of DivIVAEf mutations failed to induce such alterations.

Key Words: Enterococcus faecalis, cell division, DivIVA, mutagenesis, protein interactions

§ Current Address: Canadian Blood Services 1800 Alta Vista Drive, Ottawa, Ontario, Canada K1G 4J5


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.