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DNA Repair and Mutagenesis

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Page 1: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of

DNA Repair and Mutagenesis

Page 2: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 3: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 4: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 5: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 6: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 7: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 8: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 9: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of

Main sites of modification of DNA bases by methylating agents.

The arrow sizes reflect the relative yields of the different methylated products. Atoms or groups involved in Watson–Crick hydrogen bonding are shown in red. Bases methylated at positions indicated by the black arrows are processed primarily by AlkA; those methylated at sites marked with blue arrows are addressed by O6-methylguanine DNA methyltransferase, and those methylated at positions marked with the green arrows are processed by AlkB

Page 10: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 11: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of

AlkB-mediated demethylation of N3-methyladenine and N1-methylcytosine.

The methyl groups are converted to hydroxymethyl moieties, which are spontaneously lost in the form of formaldehyde to regenerate the unmodified bases (the aberrant methyl groups and their oxidation products are shown in red). Note that the cofactors (blue) are consumed during the reaction in stoichiometric amount

Page 12: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 13: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 14: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 15: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 16: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 17: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 18: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 19: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 20: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 21: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 22: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of

WT uvrA uvrB uvrCUV dose J/m2

05

10204080

120

E. coli that are unable to remove DNA lesions are sensitive to UV irradiation

Page 23: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of

People that are unable to remove DNA lesions are also sensitive to UV irradiation

Photo taken from Friedberg, EC., Walker GC., Siede, W (1995) in DNA Repair and Mutagenesis (ASM Press, Washington DC)

Page 24: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 25: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 26: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 27: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 28: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of

OFFOFFOFFOFFONONONONumuCuvrArecAlexAOFFOFFOFFOFFumuCuvrArecAlexAOFFOFFOFFOFFumuCuvrArecAlexAumuCuvrArecAlexAumuCuvrArecAOFFOFFOFFOFFlexALexA normally binds to the promoter regions and represses more than 40 genes

However, following UV-induced DNA damage or other replication arresting events...

...RecA binds to the single strand regions that are generated when replication forks are arrested

The bound RecA serves to protect the replication fork DNA from degradation and also promotes the autocatalyic cleavage and subsequent degradation of the LexA protein, leading to the derepression of the SOS genes.

Once the arresting lesions have been repaired and replication has been restored, the single strand substrate for RecA binding is no longer present, LexA is no longer cleaved and its cellular concentration accumulates. Then, once again, LexA binds to the promoters and represses the expression of the SOS genes, including that of lexA and recA.

=RecA=LexA=UV-induced DNA damage DNA replication bubble

Page 29: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 30: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 31: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 32: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of

0.1

1

10

oraA

0 20 40 60minutes

recA

0 20 40 60minutes

lexA-77

2820 kb

-UV

lexA1

+UV

wildtype

Page 33: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 34: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
Page 35: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of
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Page 37: DNA Repair and Mutagenesis. Main sites of modification of DNA bases by methylating agents. The arrow sizes reflect the relative yields of

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