2/15/2023 0 Comments Kaleidagraph 3.6 free download![]() Because pol κ was able to bypass these ICLs, biological evidence for a role for pol κ in tolerating the N 2- N 2-guanine ICLs was sought both cell survival and chromosomal stability were adversely affected in pol κ-depleted cells following mitomycin C exposure. Further analyses showed that although yeast Rev1 could incorporate a dCTP opposite the cross-linked guanine, no evidence was found for TLS by pol ζ or a pol ζ/Rev1 combination. The efficiency of TLS was greatly enhanced by truncation of both the 5 ′ and 3 ′ ends of the nontemplating strand. Using a series of oligodeoxynucleotides that mimic potential intermediates in ICL repair, we demonstrate that human polymerase (pol) κ not only catalyzed accurate incorporation opposite the cross-linked guanine but also replicated beyond the lesion, thus providing the first biochemical evidence for TLS past an ICL. Here, data are presented that mammalian cell replication of DNAs containing these lesions was ∼97% accurate. To identify DNA polymerases that may function in TLS past ICLs, oligodeoxynucleotides were synthesized containing site-specific ICLs in which the linkage was between N 2-guanines, similar to cross-links formed by mitomycin C and enals. Although there exists compelling genetic evidence for a homologous recombination-independent pathway for repair of interstrand cross-links (ICLs) involving translesion synthesis (TLS), biochemical support for this model is lacking. ![]()
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