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phosphorylated forms) involved in the process of
DSBs detection and repair, as well as to collect the
information concerning the kinetics parameters of
the model, such as time of deactivation of the
specific protein. Moreover, the analysis of the
expression profile of genes involved in HR pathway
will be performed with a method of quantitative
real-time PCR in stress conditions.
The analysis of the level of double strand breaks
of DNA after treatment with specific cytotoxins will
be performed with microscope analysis of H2AX
foci assay. The amount of foci reflects the amount of
DNA breaks. The analysis of the total number of
DSBs and SSBs will be performed with comet assay.
The experiments will be performed on
mammalian cancer and normal cell lines with active
or inactive forms of the proteins involved in HR.
The whole methodology is still under development.
7 EXPECTED OUTCOME
The experimental-based mathematical model
(stochastic and deterministic) of HR repair pathway
will be presented. The HR model, together with
ATM and ATR models developed by our group, will
create a comprehensive mathematical model
describing the dynamics of the interactions
occurring in the cell from the inception of DNA
damages to making the decisions about cell fate: to
direct cell to repair by HR, to arrest cell cycle or
direct cell to apoptosis pathway during HR or before
the process of repair.
The expected outcome is an identification of at
least one new component of the HR pathway which
can be even protein or miRNA, and the parameters
of the model for most of the pathway components.
Moreover, it is expected that the developed model
will reflect the experimental data and will be a good
tool for simulation of the cell behavior during HR.
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