Characterization of genomic BCR-ABL breakpoints in chronic myeloid leukaemia by PCR.
In order to understand better the mechanism of translocation between the BCR and ABL genes in CML, we have exploited a 'bubble PCR' technique to clone genomic breakpoints. BCR-ABL junction fragments were successfully amplified and sequenced in 14/32 (43%) patients tested. Breakpoints were dispersed throughout the major breakpoint cluster region without any clustering or hot spots. In three cases Alu sequences were found at or near the breakpoint on the ABL side of the translocation but no other obvious sequence homologies were found either in BCR or ABL. The translocation event was characterized further in three other patients by amplifying the reciprocal ABL-BCR junction on the 9q+ chromosome and also normal ABL around breakpoints. In two of these patients a few nucleotides of BCR and ABL were either duplicated or deleted on translocation, suggesting that staggered cuts had been made in the DNA strand prior to recombination. In the third patient 50 bp of ABL was deleted and 159 bp of M-BCR including exon b3 was duplicated, indicating either that the single-stranded cuts may span a larger distance than previously thought or that another mechanism, perhaps involving gene conversion, may be involved in this instance.
Zhang
J G
JG
LRF Leukaemia Unit, Royal Postgraduate Medical School, Hammersmith Hospital, London.
Goldman
J M
JM
Cross
N C
NC
D001483:Base Sequence; D002892:Chromosomes, Human, Pair 22; D002899:Chromosomes, Human, Pair 9; D016044:Fusion Proteins, bcr-abl / Q000235:genetics*; D016313:Genes, abl; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D014178:Translocation, Genetic
D016044:Fusion Proteins, bcr-abl