title: Biased short tract repair of palindromic loop mismatches in mammalian cells.
abstract: Mismatch repair of palindromic loops in the presence or absence of single-base mismatches was investigated in wild-type and mismatch-binding defective mutant Chinese hamster ovary cells. Recombination intermediates with a maximum heteroduplex DNA (hDNA) region of 697 bp contained a centrally located, phenotypically silent 12-base palindromic loop mismatch, and/or five single-base mismatches. In wild-type cells, both loops and single-base mismatches were efficiently repaired (80-100%). When no other mismatches were present in hDNA, loops were retained with a 1.6-1.9:1 bias. However, this bias was eliminated when single-base mismatches were present, perhaps because single-base mismatches signal nick-directed repair. In the multiple marker crosses, most repair tracts were long and continuous, with preferential loss of markers in cis to proximal nicks, consistent with nicks directing most repair in this situation. However, approximately 25% of repair tracts were discontinuous as a result of loop-specific repair, or from segregation or short tract repair of single-base mismatches. In mutant cells, single-base mismatches were repaired less frequently, but the loop was still repaired efficiently and with bias toward loop retention, indicating that the defect in these cells does not affect loop-specific repair. Repair tracts in products from mutant cells showed a wide variety of mosaic patterns reflecting short regions of repair and segregation consistent with reduced nick-directed repair. In mutant cells, single-base mismatches were repaired more efficiently in the presence of the loop than in its absence, a likely consequence of corepair initiated at the loop.
authors.affiliation: Department of Cancer Biology, Harvard University School of Public Health, Boston, Massachusetts 02115, USA.
authors.forename: D G
authors.initials: DG
authors.lastname: Taghian
authors.forename: H
authors.initials: H
authors.lastname: Hough
authors.forename: J A
authors.initials: JA
authors.lastname: Nickoloff
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D004260:DNA Repair; D008322:Mammals; D016297:Mutagenesis, Site-Directed; D011995:Recombination, Genetic; D012107:Research Design