DNA-mediated redox signaling by UvrC

Abstract

The E. coli UvrC protein is an integral part of the DNA nucleotide excision repair pathway. Our lab. has focused on studies of redox DNA signaling among DNA repair proteins contg. 4Fe- 4S clusters. We have now explored whether UvrC may also contain a 4Fe- 4S cluster. A new purifn. route was developed to express an MBP- UvrC fusion. The purified protein shows a UVvisible absorbance spectrum consistent with that of a 4Fe- 4S cluster. ESR expts. of reduced UvrC give rise to a signal indicative of a 4Fe- 4S cluster. Electrochem. expts. indicate that the protein is electrochem. active on DNA- modified gold electrodes. Finally, genetic methods to study possible DNA- mediated signaling between UvrC and other DNA- processing enzymes contg. 4Fe- 4S clusters are being examd.

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