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Molecular characterization of the genomic breakpoint junction in a t(11;22) translocation in Ewing sarcoma.
Genes Chromosomes Cancer. 1999 May; 25(1):6-15.GC

Abstract

Polymerase chain reaction (PCR)-based nucleotide sequence analysis was performed in 12 cases of Ewing sarcoma on the cDNA and/or genomic DNA breakpoint regions of a t(11;22)(q24;q12), which joins the EWS gene located on chromosome 22 with the FLI1 gene located on chromosome 11, in order to understand the molecular mechanism of this translocation. Reverse transcriptase-PCR on total tumor cell RNA from the examined cases showed five types of EWS-FLI1 chimeric product, resulting from various junctions between EWS exon 7 or 10 with FLI1 exon 5, 6, or 8. Sequencing of the genomic fusion junctions of EWS-FLI1 in seven cases showing three types of the chimeric cDNA products revealed that most of the breakpoint junctions shared common nucleotide(s) from both genes, and that the breakpoints in EWS introns 7 and 10 clustered within 100 bp and 300 bp, respectively. All the junctions were found to be flanked by various oligomers, among which a consensus sequence, 5'-AGAAAARDRR-3', was found near the breakpoints of both genes in four cases, suggesting that these oligomers may have a functional significance in the genesis of t(11;22). In addition to these oligomers, sequences highly homologous to Alu repeats and/or eukaryotic topoisomerase II cleavage sites were located near, or flanked, or even encompassed, the breakpoints in most of the cases examined. Thus, these sequences may also mediate DNA double-strand breakage and rejoining to generate the t(11;22). Genomic sequence analysis of both EWS-FLI1 and FLI1-EWS chimeric genes in three of the seven cases demonstrated a deletion and duplication of both EWS and FLI1 sequences in two cases and no gain or loss in one case. The present findings suggest that multiple mechanisms may be operative for the break and rejoining of the fragments of chromosomes 11 and 22 in the genesis of t(11;22), and that some of these translocations are asymmetric at the molecular level.

Authors+Show Affiliations

Laboratory of Cytogenetics, Division of Bioscience, Graduate School of Environmental Earth Science, Hokkaido University, Sapporo, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

10221334

Citation

Obata, K, et al. "Molecular Characterization of the Genomic Breakpoint Junction in a T(11;22) Translocation in Ewing Sarcoma." Genes, Chromosomes & Cancer, vol. 25, no. 1, 1999, pp. 6-15.
Obata K, Hiraga H, Nojima T, et al. Molecular characterization of the genomic breakpoint junction in a t(11;22) translocation in Ewing sarcoma. Genes Chromosomes Cancer. 1999;25(1):6-15.
Obata, K., Hiraga, H., Nojima, T., Yoshida, M. C., & Abe, S. (1999). Molecular characterization of the genomic breakpoint junction in a t(11;22) translocation in Ewing sarcoma. Genes, Chromosomes & Cancer, 25(1), 6-15.
Obata K, et al. Molecular Characterization of the Genomic Breakpoint Junction in a T(11;22) Translocation in Ewing Sarcoma. Genes Chromosomes Cancer. 1999;25(1):6-15. PubMed PMID: 10221334.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Molecular characterization of the genomic breakpoint junction in a t(11;22) translocation in Ewing sarcoma. AU - Obata,K, AU - Hiraga,H, AU - Nojima,T, AU - Yoshida,M C, AU - Abe,S, PY - 1999/4/30/pubmed PY - 2000/6/20/medline PY - 1999/4/30/entrez SP - 6 EP - 15 JF - Genes, chromosomes & cancer JO - Genes Chromosomes Cancer VL - 25 IS - 1 N2 - Polymerase chain reaction (PCR)-based nucleotide sequence analysis was performed in 12 cases of Ewing sarcoma on the cDNA and/or genomic DNA breakpoint regions of a t(11;22)(q24;q12), which joins the EWS gene located on chromosome 22 with the FLI1 gene located on chromosome 11, in order to understand the molecular mechanism of this translocation. Reverse transcriptase-PCR on total tumor cell RNA from the examined cases showed five types of EWS-FLI1 chimeric product, resulting from various junctions between EWS exon 7 or 10 with FLI1 exon 5, 6, or 8. Sequencing of the genomic fusion junctions of EWS-FLI1 in seven cases showing three types of the chimeric cDNA products revealed that most of the breakpoint junctions shared common nucleotide(s) from both genes, and that the breakpoints in EWS introns 7 and 10 clustered within 100 bp and 300 bp, respectively. All the junctions were found to be flanked by various oligomers, among which a consensus sequence, 5'-AGAAAARDRR-3', was found near the breakpoints of both genes in four cases, suggesting that these oligomers may have a functional significance in the genesis of t(11;22). In addition to these oligomers, sequences highly homologous to Alu repeats and/or eukaryotic topoisomerase II cleavage sites were located near, or flanked, or even encompassed, the breakpoints in most of the cases examined. Thus, these sequences may also mediate DNA double-strand breakage and rejoining to generate the t(11;22). Genomic sequence analysis of both EWS-FLI1 and FLI1-EWS chimeric genes in three of the seven cases demonstrated a deletion and duplication of both EWS and FLI1 sequences in two cases and no gain or loss in one case. The present findings suggest that multiple mechanisms may be operative for the break and rejoining of the fragments of chromosomes 11 and 22 in the genesis of t(11;22), and that some of these translocations are asymmetric at the molecular level. SN - 1045-2257 UR - https://www.unboundmedicine.com/medline/citation/10221334/Molecular_characterization_of_the_genomic_breakpoint_junction_in_a_t_11 L2 - https://onlinelibrary.wiley.com/resolve/openurl?genre=article&sid=nlm:pubmed&issn=1045-2257&date=1999&volume=25&issue=1&spage=6 DB - PRIME DP - Unbound Medicine ER -