TY - JOUR
T1 - Purification and Characterization of Escherichia coli DNA Polymerase V
AU - Schlacher, Katharina
AU - Jiang, Qingfei
AU - Woodgate, Roger
AU - Goodman, Myron F.
N1 - Funding Information:
This work was supported by NIH grants ESO12259 and R37GM21422 to MFG and by the NIH Intramural Research Program to RW.
PY - 2006
Y1 - 2006
N2 - Cell survival and genome rescue after UV irradiation in Escherichia coli depends on DNA repair mechanisms induced in response to DNA damage as part of the SOS regulon. SOS occurs in two phases. The first phase is dominated by accurate repair processes such as excision and recombinational DNA repair, while the second phase is characterized by a large ∼100-fold increase in mutations caused by an error-prone replication of damaged DNA templates. SOS mutagenesis occurs as a direct result of the action of the UmuDC gene-products, which form the low fidelity Escherichia coli DNA polymerase V, a heterotrimeric complex composed of UmuD′2C. This chapter describes the preparation of highly purified native pol V that is suitable for a wide range of biochemical studies of protein-protein, protein-DNA interactions and translesion-synthesis (TLS) mechanisms.
AB - Cell survival and genome rescue after UV irradiation in Escherichia coli depends on DNA repair mechanisms induced in response to DNA damage as part of the SOS regulon. SOS occurs in two phases. The first phase is dominated by accurate repair processes such as excision and recombinational DNA repair, while the second phase is characterized by a large ∼100-fold increase in mutations caused by an error-prone replication of damaged DNA templates. SOS mutagenesis occurs as a direct result of the action of the UmuDC gene-products, which form the low fidelity Escherichia coli DNA polymerase V, a heterotrimeric complex composed of UmuD′2C. This chapter describes the preparation of highly purified native pol V that is suitable for a wide range of biochemical studies of protein-protein, protein-DNA interactions and translesion-synthesis (TLS) mechanisms.
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U2 - 10.1016/S0076-6879(06)08023-2
DO - 10.1016/S0076-6879(06)08023-2
M3 - Review article
C2 - 16793381
AN - SCOPUS:33745188227
SN - 0076-6879
VL - 408
SP - 378
EP - 390
JO - Methods in enzymology
JF - Methods in enzymology
ER -