Source: http://www.asmscience.org/content/book/10.1128/9781555816704.ch17
Timestamp: 2019-04-25 18:13:07+00:00

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Detection of untargeted mutations in a budding-yeast mating system. UV-irradiated haploid cells containing a CYC1 deletion (cyc1Δ-363) are fused with a nonirradiated partner carrying a point mutation (cyc1-91) in the region deleted in cyc1Δ-363. Mutants with reversion of the point mutation (cyc1-91 → CYC+) in the unirradiated genome are detected among the diploid progeny.
ade2 adeX mutational system for determining the timing of mutation fixation in S. cerevisiae. (A) ade2 mutants form red colonies as a result of the accumulation of an intermediate of the adenine biosynthetic pathway. Inactivation of a metabolic step upstream of ade2 by a second mutation (adeX) prevents the formation of the red pigment; hence, the colony is white. (B) If (stationary-phase haploid) ade2 cells are UV irradiated and an adeX mutation has undergone mutational fixation in both DNA strands prereplicatively (lower section), a pure-white colony will result. If the sequence alteration has affected only one DNA strand before replication, one daughter cell will show the ade2-conferred phenotype and (if this cell remains viable) a red-and-white sectored colony will result (upper section).
Alkaline sucrose density gradient profiles of DNA from unirradiated or X-irradiated (10 Gy) Chinese hamster ovary (CHO) cells. In the irradiated cells there is a decrease in the relative amount of small DNA molecules (fractions 5 to 10), reflecting regions in which replication was initiated soon after irradiation. However, the relative number of large molecules (fractions 15 to 20), which reflect the progression of replicons that were initiated prior to irradiation, is not significantly decreased. This suggests that growing DNA strands continue to be elongated but new chains cannot be initiated.
Schematic view of one-step and two-step DNA lesion bypass. A bypass polymerase replaces a high-fidelity polymerase that cannot replicate past a lesion in the template. TLS is accomplished in a single step (left) or in two separate steps (right). The latter mechanism involves base insertion opposite a lesion and elongation that is carried out by a separate polymerase specialized in primer extension. Both steps can be error-prone. The low processivity of these translesion polymerases facilitates replacement by a replicative polymerase a short distance downstream of the lesion.
Subpathways of replicative DNA damage tolerance defined by the proteins of the S. cerevisiae Rad6 group. Rad6 functions as an E2 enzyme in Ubr1-mediated, N-end-rule-dependent protein degradation, independently of DNA damage tolerance pathways. In DNA damage responses, complex formation of Rad6 with the single-stranded DNA-binding Rad18 protein is essential. Both proteins are required for all error-free and error-prone pathways, and cross talk with proteins mediating TLS must be assumed. Proteins of the error-free subpathway are being attracted to sites of stalled replication through the Rad18-Rad5-Ubc13 interaction. Ubc13-Mms2 represents another ubiquitin (Ub)-conjugating machinery that attaches Lys63-linked ubiquitin residues to unknown (?) substrates.
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