Immunological detection of UV induced cyclobutane pyrimidine dimers and (6-4) photoproducts in DNA from reference bacteria and natural aquatic populations.
UV light-caused DNA damage is a widespread field of study. As UV light has the biological effect of inactivating or killing bacteria, it is used for water disinfection. Due to this application, it is important to study the DNA damage efficiencies and regeneration capacities in bacteria after UV irradiation. Two monoclonal antibodies, anti-CPD and anti-(6-4) PP, were applied for an immunoassay of UV-induced DNA modifications. Cyclobutane pyrimidine dimer (CPD) and 6-4 photoproduct (6-4 PP) were detected in the reference bacteria Pseudomonas aeruginosa and Enterococcus faecium, and in natural water communities. The antibody-mediated detection signals increased with the UV doses from 100-400J/m(2). Here, the CPD-specific signals were stronger than the (6-4) PP-specific signals. These immunological results were in accordance with parallel cultivation experiments. All UV-irradiated bacteria showed a reduction of their growth rate depending on UV application by several orders of magnitudes. The immunoassay was also applied to three types of natural aquatic habitats with different cell densities. Besides artificial UV irradiation, it was possible to visualize natural sunlight effects on these natural bacterial communities. Light-dependent and dark repair processes were distinguished using the established immunological assays. The antibody-mediated analyses presented are fast methods to detect UV-induced DNA lesions and repair capacities in selected bacterial species as well as in entire natural mixed populations.
Karlsruhe Institute of Technology (KIT)-Campus North, Institute of Functional Interfaces (IFG), Microbiology of Natural and Technical Interfaces Department, P.O. Box 3640, D-76021 Karlsruhe, Germany.
Stephanie
S
Kraft
Kraft
K
Stephanie
Ursula
U
Obst
Obst
O
Ursula
Thomas
T
Schwartz
Schwartz
S
Thomas
D001419:Bacteria / Q000528:radiation effects*; D004247:DNA / Q000528:radiation effects*; D004249:DNA Damage; D007118:Immunoassay / Q000379:methods; D050296:Microbial Viability / Q000528:radiation effects; D011740:Pyrimidine Dimers / Q000032:analysis*; D014466:Ultraviolet Rays; D014871:Water Microbiology
D011740:Pyrimidine Dimers; D004247:DNA