Enterococcus faecalis divIVA: an essential gene involved in cell division, cell growth and chromosome segregation

• Published in: Microbiology (Society for General Microbiology) 2005-05, Vol.151 (5), p.1381-1393
• Main Authors: Ramirez-Arcos, Sandra, Liao, Mingmin, Marthaler, Susan, Rigden, Marc, Dillon, Jo-Anne R
• Format: Article
• Language: English
• Subjects: Bacillus subtilis; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Cell Cycle Proteins - genetics; Cell Cycle Proteins - metabolism; Cell Division; Chromosome Segregation; Enterococcus; Enterococcus faecalis; Enterococcus faecalis - cytology; Enterococcus faecalis - genetics; Enterococcus faecalis - growth & development; Enterococcus faecalis - metabolism; Escherichia coli; Escherichia coli - cytology; Escherichia coli - genetics; Escherichia coli - growth & development; Escherichia coli - metabolism; Gene Deletion; Genes, Essential; Molecular Sequence Data; Sequence Analysis, DNA; Species Specificity; Streptococcus; Streptococcus pneumoniae
• ISSN: 1350-0872; 1465-2080
• DOI: 10.1099/mic.0.27718-0

1 Department of Biochemistry, Microbiology and Immunology, University of Ottawa, Ottawa, ON, Canada K1H 8M5 2 Centre for Research in Biopharmaceuticals and Biotechnology, University of Ottawa, Ottawa, ON, Canada K1H 8M5 Correspondence Jo-Anne R. Dillon j.dillon{at}usask.ca Enterococcus faecalis divIVA ( divIVA Ef ) is an essential gene implicated in cell division and chromosome segregation. This gene was disrupted by insertional inactivation creating E. faecalis JHSR1, which was viable only when a wild-type copy of divIVA Ef was expressed in trans , confirming the essentiality of the gene. The absence of DivIVA Ef in E. faecalis JHSR1 inhibited proper cell division, which resulted in abnormal cell clusters possessing enlarged cells of altered shape instead of the characteristic diplococcal morphology of enterococci. The lower viability of the divIVA Ef mutant is caused by improper nucleoid segregation and impaired septation within the numerous cells generated in each cluster. Overexpression of DivIVA Ef in Escherichia coli KJB24 resulted in enlarged cells with disrupted cell division, suggesting that this round E. coli mutant strain could be used as an indicator for functionality of DivIVA Ef . A Bacillus subtilis divIVA mutant was not complemented by DivIVA Ef , indicating that this protein does not recognize DivIVA-specific target sites in B. subtilis , or that it does not interact with other proteins of the cell division machinery of this micro-organism. DivIVA Ef also failed to complement a Streptococcus pneumoniae divIVA mutant, supporting the phylogenetic distance between Enterococcus and Streptococcus . Our results indicate that DivIVA is a species-specific multifunctional protein implicated in cell division and chromosome segregation in E. faecalis . Abbreviations: Amp, ampicillin; Cm, chloramphenicol; Ery, erythromycin; Kan, kanamycin The GenBank/EMBL/DDBJ accession number for the sequence reported in this paper is AF414352 . Present address: Infectious Diseases, Canadian Blood Services, 1800 Alta Vista Drive, Ottawa, Ontario K1G 4J5, Canada. Present address: College of Arts and Science, University of Saskatchewan, Room 226, Arts Building, 9 Campus Drive, Saskatoon, Saskatchewan, Canada S7N 5A5.