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Blood, Vol. 107, Issue 2, 777-780, January 15, 2006
A model of oncogenic rearrangements: differences between chromosomal translocation mechanisms and simple double-strand break repair
Blood Weinstock et al.
107: 777
Supplemental materials for: Weinstock et al
Files in this Data Supplement:
- Supplemental Methods (PDF, 105 KB)
- Table S1A. Summary of deletions and duplications to the parental chromosomes in patient-derived translocation junctions compared with those derived in the current study (PDF, 65.3 KB) -
As the precise breakpoint is unknown for most cancer-associated translocations, the total deletion or duplication length is derived from the parental chromosomes participating in the reciprocal translocation rather than from the derivative chromosomes. For comparison, data from the neo+ clones derived in the current study is compiled in a similar manner from the parental chromosomes. (Note that the data presented in Figures 2 and 3 are derived from the derivative chromosomes, not the parental chromosomes.)
- Table S1B. Summary of insertions and microhomology use in the derivative chromosomes of patient-derived translocation junctions compared with those derived from the current study (PDF, 87.1 KB)
- Figure S1. Structure of translocation junctions containing complex events and large insertions (PDF, 334 KB) -
Der(17) in neo+ clones T7 and T37, which are derived from the p5rE reporter cell line 18, contain locally-derived inverted sequences and duplicated sequences in a direct orientation, as well as non-templated nucleotide insertions (blue). Der(14) in neo+ clones T21 and T22, which are derived from the pCr15 cell line, contain duplicated sequences in a direct orientation and non-templated nucleotide insertions. Interestingly, in clone T22 the duplicated sequence is derived from the centromeric side of the chr.17 breakpoint, rather than the telomeric side that contributes to the der(14) junction. Large insertions are derived from various sources, as indicated. The top strand of the chromosome ends at the I-SceI site are shown for each of the translocation reporters prior to NHEJ with the I-SceI overhangs filled-in. The filled-in 3' overhang is apparent in the complex junction from clone pCr15-T22 (underline).
- Figure S2 (PDF, 94.9 KB) -
A. Comparison of p5rE der(17) junctions after translocation formation and intrachromosomal repair upon I-SceI expression. Translocation junctions are derived from two parental cell lines (p5rE18 and p5rE26) after neo+ selection of individual clones. Not shown are the 13 translocation junctions that reestablished the I-SceI site, which are identical to the der(17) sequence (top gray box). Additionally, two complex translocation junctions and a junction containing an insertion are diagrammed in Figure S1. Intrachromosomal junctions are derived from two translocation clones (p5rE18-T1 and p5rE26-T3) using the strategy shown in Fig. 2D to obtain plasmid clones. The number of cell clones (translocations) and plasmid clones (intrachromosomal repair) with an identical junction sequence are indicated in parentheses. Since translocation junctions are derived from neo+ clonal analysis, junctions that occur repetitively are derived from independent events, often being derived from different parental cell lines or different experiments of the same cell line, or if they are from the same experiment, the der(14) junction differs, reinforcing the independence of the events. (e.g., the 7 junctions labeled p5rE18-T3). For the intrachromosomal junctions, those that occur repetitively are also often obtained from the different cells or experiments, as indicated by the slash marks in the parentheses. B. Sequences of p5rErev and pCr15 der(17) and p5rE, p5rErev and pCr15 der(14) junctions that repaired by NHEJ. Complex translocation junctions are diagrammed in Supplementary Fig. 1. Der(14) junctions that repaired by SSA (n=62) and junctions confirmed by Southern blotting but not sequenced (n=10) are not included. Microhomology, underlined; inserted nucleotides, red; modifications away from the ends, gray boxes; NA, not applicable.
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