Characterisation of the localisation and function of the Bacillus subtilis YuaB protein during biofilm formation

Bacteria can actively communicate and interact with each other to establish multicellular communities. Many of these processes involve functional differentiation of cells into specialised subpopulations by expression of varying genetic programmes. This leads to division of labour between the arising subpopulations of cells in the community. One type of such community is the biofilm, which is composed of microbial cells enclosed in a biopolymeric matrix. Such biofilms can be formed in a large range of environments from sea beds to animal tissues. Bacillus subtilis is a soil dwelling Gram-positive rod that was shown to closely interact with plants and establish a protective symbiosis by formation of biofilms on the roots. The biofilm matrix synthesised by B. subtilis is composed of the exopolysaccharide, for which the chemical structure is not yet established, and a protein TasA that forms amyloid-like fibres spanning between the cells and anchored to the cells by an accessory protein TapA. A third protein of unknown function, YuaB, has also been shown to be necessary for establishment of a biofilm. However, the mechanism of function for YuaB has not been elucidated. The data presented in this report focus on the role played by YuaB during formation of the biofilm. By analysis of cell differentiation patterns YuaB was shown to be required for maturation of the biofilm. The localisation of YuaB is identified in two “subtypes” of biofilm, a biofilm pellicle floating on the air-liquid interface and complex colonies formed on solid surfaces. This is achieved using a combination of biofilm fractionation combined with Western blotting and a newly developed method for immuno-fluorescent labelling of biofilm proteins. YuaB acts in synergy with the exopolysaccharide and TasA, as both components of the biofilm matrix are synthesised in the absence of YuaB but the biofilm is not made. The initial structural characterisation of YuaB is presented based on in silico predictions and physiological and biophysical analysis of the mutations introduced into the sequence of YuaB. The experimental data is concluded with a hypothesis that YuaB forms a hydrophobic protective layer necessary for support of the structure of the matured biofilm.

Name dc:type.qualificationname

Doctor of Philosophy

Level dc:type.qualificationlevel

Doctoral Thesis

Grantor dc:publisher.institution

University of Dundee

Year dc:date.issued

2012

Author dc:creator

• Ostrowski, Adam

Advisor dc:contributor.advisor

• Stanley-Wall, Nicola

• Bacillus subtilis
• YuaB
• Biofilms
• Biofilm formation
• BslA

Language dc:language

eng

Identifier

oai:discovery.dundee.ac.uk:studenttheses/29fe0d1b-a010-4cb9-8698-7f393b6e08ee

OAI identifier oai:identifier

oai:discovery.dundee.ac.uk:studenttheses/29fe0d1b-a010-4cb9-8698-7f393b6e08ee