3FNC1†Present address: Infectious Diseases, Canadian Blood Services, 1800 Alta Vista Drive, Ottawa, Ontario K1G 4J5, Canada.
3FNC2‡Present address: College of Arts and Science, University of Saskatchewan, Room 226, Arts Building, 9 Campus Drive, Saskatoon, Saskatchewan, Canada S7N 5A5.
Enterococcus faecalis divIVA (divIVAEf) 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 divIVAEf was expressed in trans, confirming the essentiality of the gene. The absence of DivIVAEf 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 divIVAEf mutant is caused by improper nucleoid segregation and impaired septation within the numerous cells generated in each cluster. Overexpression of DivIVAEf 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 DivIVAEf. A Bacillus subtilis divIVA mutant was not complemented by DivIVAEf, 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. DivIVAEf 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.
AkiyamaT.,
InouyeS.,
KomanoT.
2003; Novel developmental genes, fruCD, of Myxococcus xanthus: involvement of a cell division protein in multicellular development. J Bacteriol 185:3317–3324[CrossRef]
BaeT.,
Clerc-BardinS.,
DunnyG. M.
2000; Analysis of expression of prgX, a key negative regulator of the transfer of the Enterococcus faecalis pheromone-inducible plasmid pCF10. J Mol Biol 297:861–875[CrossRef]
BradfordM. M.
1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254[CrossRef]
BryanE. M.,
BaeT.,
KleerebezemM.,
DunnyG. M.
2000; Improved vectors for nisin-controlled expression in gram-positive bacteria. Plasmid 44:183–190[CrossRef]
CalleganM. C.,
JettB. D.,
HancockL. E.,
GilmoreM. S.
1999; Role of hemolysin BL in the pathogenesis of extraintestinal Bacillus cereus infection assessed in an endophthalmitis model. Infect Immun 67:3357–3366
ChristieP. J.,
DunnyG. M.
1986; Identification of regions of the Streptococcus faecalis plasmid pCF-10 that encode antibiotic resistance and pheromone response functions. Plasmid 15:230–241[CrossRef]
CorbinB. D.,
YuX. C.,
MargolinW.
2002; Exploring intracellular space: function of the Min system in round-shaped Escherichia coli. EMBO J 21:1998–2008[CrossRef]
de BoerP. A.,
CrossleyR. E.,
RothfieldL. I.
1989; A division inhibitor and a topological specificity factor coded for by the minicell locus determine proper placement of the division septum in E. coli
. Cell 56:641–649[CrossRef]
de la FuenteA.,
PalaciosP.,
VicenteM.
2001; Transcription of the Escherichia coli dcw cluster: evidence for distal upstream transcripts being involved in the expression of the downstream ftsZ gene. Biochimie 83:109–115[CrossRef]
EdwardsD. H.,
ErringtonJ.
1997; The Bacillus subtilis DivIVA protein targets to the division septum and controls the site specificity of cell division. Mol Microbiol 24:905–915[CrossRef]
FacklamR. R.,
CarvalhoM. G. S.,
TeixeiraL. M.
2002; History, taxonomy, biochemical characteristics, and antibiotic susceptibility testing of enterococci. In The Enterococci, Pathogenesis, Molecular Biology, and Antibiotic Resistance pp 1–54 Edited by
GilmoreM. S.,
ClewellD. B.,
CourvalinP.,
DunnyG. M.,
MurrayB. E.,
RiceL. B.
Washington, DC: American Society for Microbiology;
FaddaD.,
PischeddaC.,
CaldaraF.,
WhalenM. B.,
AnderluzziD.,
DomeniciE.,
MassiddaO.
2003; Characterization of divIVA and other genes located in the chromosomal region downstream of thedcw cluster in Streptococcus pneumoniae
. J Bacteriol 185:6209–6214[CrossRef]
HigginsM. L.,
Daneo-MooreL.,
BoothbyD.,
ShockmanG. D.
1974; Effect of inhibition of deoxyribonucleic acid and protein synthesis on the direction of cell wall growth in Streptococcus faecalis
. J Bacteriol 118:681–692
HigginsM. L.,
CarsonD. D.,
Daneo-MooreL.
1980; Morphological effect of cerulenin treatment on Streptococcus faecalis as studied by ultrastructure reconstruction. J Bacteriol 143:989–994
JacobA. E.,
HobbsS. J.
1974; Conjugal transfer of plasmid-borne multiple antibiotic resistance in Streptococcus faecalis var.zymogenes
. J Bacteriol 117:360–372
JonesL. J.,
Carballido-LopezR.,
ErringtonJ.
2001; Control of cell shape in bacteria: helical, actin-like filaments in Bacillus subtilis. Cell 104:913–922[CrossRef]
MarstonA. L.,
ErringtonJ.
1999; Selection of the midcell division site in Bacillus subtilis through MinD-dependent polar localization and activation of MinC. Mol Microbiol 33:84–96[CrossRef]
MassiddaO.,
AnderluzziD.,
FriedliL.,
FegerG.
1998; Unconventional organization of the division and cell wall cluster of Streptococcus pneumoniae. Microbiology 144:3069–3078[CrossRef]
PoyartC.,
Trieu-CuotP.
1997; A broad-host-range mobilizable shuttle vector for the construction of transcriptional fusions to beta-galactosidase in gram-positive bacteria. FEMS Microbiol Lett 156:193–198[CrossRef]
PucciM. J.,
ThanassiJ. A.,
DiscottoL. F.,
KesslerR. E.,
DoughertyT. J.
1997; Identification and characterization of cell-wall division gene clusters in pathogenic Gram positive cocci. J Bacteriol 179:5632–5635
Ramirez-ArcosS.,
SzetoJ.,
DillonJ. A.,
MargolinW.
2002; Conservation of dynamic localization among MinD and MinE orthologues: oscillation of Neisseria gonorrhoeae proteins in Escherichia coli
. Mol Microbiol 46:493–504[CrossRef]
Ramirez-ArcosS.,
GrecoV.,
DouglasH.,
TessierD.,
FanD.,
SzetoJ.,
WangJ.,
DillonJ. R.
2004; Conserved glycines in the C-terminus of MinC proteins are implicated in their functionality as a cell division inhibitor. J Bacteriol 186:2841–2855[CrossRef]
RamosA.,
HonrubiaM. P.,
ValbuenaN.,
VaqueraJ.,
MateosL. M.,
GilJ. A.
2003; Involvement of DivIVA in the morphology of the rod-shaped actinomycete Brevibacterium lactofermentum. Microbiology 149:3531–3542[CrossRef]
SzetoJ.,
Ramirez-ArcosS.,
RaymondC.,
HicksL. D.,
KayC. M.,
DillonJ. A.
2001; Gonococcal MinD affects cell division in Neisseria gonorrhoeae and Escherichia coli and exhibits a novel self-interaction. J Bacteriol 183:6253–6264[CrossRef]
The National Committee for Clinical Laboratory Standards (NCCLS)2002Performance Standards for Antimicrobial Susceptibility Testing; Twelfth Informational Supplement NCCLS document M100_MS12
ThomaidesH. B.,
FreemanM.,
El KarouiM.,
ErringtonJ.
2001; Division site selection protein DivIVA of Bacillus subtilis has a second distinct function in chromosome segregation during sporulation. Genes Dev 15:1662–1673[CrossRef]
WuL. J.,
ErringtonJ.
2003; RacA and the Soj-Spo0J system combine to effect polar chromosome segregation in sporulating Bacillus subtilis. Mol Microbiol 49:1463–1475[CrossRef]
WuL. J.,
ErringtonJ.
2004; Coordination of cell division and chromosome segregation by a nucleoid occlusion protein in Bacillus subtilis. Cell 117:915–925[CrossRef]