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RT‐PCR analysis of the MOZ‐CBP and CBP‐MOZ chimeric transcripts in acute myeloid leukemias with t(8;16)(p11;p13)
Author(s) -
Panagopoulos Ioannis,
Isaksson Margareth,
Lindvall Charlotta,
Björkholm Magnus,
Ahlgren Tomas,
Fioretos Thoas,
Heim Sverre,
Mitelman Felix,
Johansson Bertil
Publication year - 2000
Publication title -
genes, chromosomes and cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.754
H-Index - 119
eISSN - 1098-2264
pISSN - 1045-2257
DOI - 10.1002/1098-2264(200008)28:4<415::aid-gcc7>3.0.co;2-i
Subject(s) - microbiology and biotechnology , fusion transcript , biology , primer (cosmetics) , southern blot , myeloid leukemia , gene , chromosomal translocation , polymerase chain reaction , reverse transcriptase , genetics , chemistry , cancer research , organic chemistry
The translocation t(8;16)(p11;p13) is associated with a subtype of acute monocytic leukemia (AML M5) characterized morphologically by erythrophagocytosis and clinically by a poor prognosis. The t(8;16) fuses the MOZ gene from 8p11 with the CBP (also named CREBBP ) gene from 16p13. Previously published studies of MOZ and CBP rearrangements in t(8;16)‐positive AML have used fluorescence in situ hybridization and Southern blot methodologies, whereas attempts to amplify and to analyze further the chimeric MOZ ‐ CBP and CBP ‐ MOZ transcripts by means of reverse transcriptase‐polymerase chain reaction (RT‐PCR) have largely been unsuccessful. In the only t(8;16) that has been described at the sequence level using RT‐PCR, the CBP ‐ MOZ fusion was found to be out‐of‐frame, suggesting that the reciprocal MOZ ‐ CBP transcript is the essential one for leukemogenesis. We have developed an RT‐PCR strategy that enables us to detect the MOZ ‐ CBP as well as the CBP ‐ MOZ fusions in the two AML M5 with t(8;16)(p11;p13) analyzed. In both leukemias, the combination of a MOZ forward and a CBP reverse primer amplified a strongly expressed 1,128 bp fragment (type I transcript) and a weakly expressed 415 bp fragment (type II transcript). In the type I transcript, nucleotide (nt) 3,745 of MOZ was fused in‐frame with nt 284 of CBP , whereas in the type II transcript, nt 3,745 of MOZ was fused out‐of‐frame with nt 997 of CBP . Nested PCR with a combination of two forward CBP and two reverse MOZ primers amplified CBP ‐ MOZ chimeric transcripts in both cases. Direct sequence analysis showed that nt 283 of CBP was fused in‐frame with nt 3,746 of MOZ , that the initiation ATG codon of the CBP gene remained intact, and that there was no mutation or deletion in the part of the CBP gene included in the CBP ‐ MOZ transcript. Thus, the data we present are not informative with regard to the question whether it is the MOZ ‐ CBP or the CBP ‐ MOZ transcript that is leukemogenic. The present RT‐PCR method may be of value for rapid identification of the t(8;16) and also for further molecular genetic studies of the two fusion transcripts and their roles in leukemogenesis. Genes Chromosomes Cancer 28:415–424, 2000. © 2000 Wiley‐Liss, Inc.