Determining the role of coiled-coil domain interactions in the oligomerization of DivIVA from Enterococcus faecalis

Abstract

The involvement of DivIVA in cell division and chromosome segregation has been described in a number of Gram positive organisms. To date, most studies have focused on the effect of overexpression or disruption of DivIVA in various bacterial backgrounds. However, no detailed biochemical study of DivIVA has been conducted to identify which domains are responsible for the different functions of DivIVA. This study examines the coiled-coil domains of DivIVA from the Gram positive coccus Enterococcus faecalis to determine their role in oligomer formation. Preliminary localization studies of DivIVAEf in both homologous and heterologous backgrounds were also performed. While no conclusive results were obtained for GFP studies, immunogold electron microscopy of DivIVA in E. faecalis showed localization to the division septum and cell poles, similar to DivIVA from other organisms. Once proper GFP localization studies can be carried out, the bank of mutations generated in this study can be used to determine the region(s) of DivIVAEf involved in division site localization. (Abstract shortened by UMI.)