Uncoiling the mysteries of DivIVA(Ef): The biological functioning of the Enterococcus faecalis cell development protein

Abstract

Cell division mechanisms involving native proteins MinCDE or MinCD and DivIVA have been explored using Esherichia coli (Ec) and Bacillus subtilis (Bs), respectively. My research examines the Gram-positive coccal-shaped Enterococcus faecalis (Ef) protein DivIVA, which functions without any Min proteins. To determine whether E. coli could be used as a suitable indicator system, DivIVAEf overexpression and morphological studies were undertaken. Constructs with unique mutations created along the 699-nucleotide length of the divIVAEf gene to abrogate specific coiled-coil domains at the N-terminal, C-terminal and two central regions were investigated in this model. The original mutations were previously used in different systems and therefore required PCR amplification, double digest with new restriction enzymes prior to subcloning in pUC18 for transformation in E. coli (Ramirez-Arcos et al., 2005; Rigden et al., 2005). (Abstract shortened by UMI.)