A mathematical model of in vitro reconstituted Base Excision repair as operative on IUdR

generated mispairs

These hybrid models will include the discrete states of the Base Excision repair process along with the underlying continuous-time dynamics that describe the fundamental biochemical reactions between the enzymes and substrates.

Breakage of the AP site

Resolution of the bands on a denaturing urea gel followed by

fluorescence scanning

G

AP site

G

IUdRMED1

dsDNA with the mispair

dsDNA with the AP site

DNA glycosylase activity

Alkali/endonuclease treatment

Untreated dsDNA

Breakage of the AP site

Resolution of the bands on a denaturing urea gel followed by

fluorescence scanning

G

AP site

G

IUdRMED1

dsDNA with the mispair

dsDNA with the AP site

DNA glycosylase activity

G

AP site

G

AP site

G

IUdRMED1

dsDNA with the mispair G

IUdRMED1

dsDNA with the mispair

dsDNA with the AP site

DNA glycosylase activity

Alkali/endonuclease treatment

Untreated dsDNA

An in vitro fast, convenient and non-radioactive assay for quantitative measurements of enzymatic activity is developed. This assay will facilitate the generation of kinetic data for feeding the hybrid model

A GFP based in vivo BER assay is developed to validate the model under native conditions

Project 1