Truncated c-Myb expression in the human leukemia cell line TK-6

Akihiro Tomita, T. Watanabe, H. Kosugi, H. Ohashi, T. Uchida, T. Kinoshita, S. Mizutani, T. Hotta, T. Murate, M. Seto, H. Saito

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

The c-MYB proto-oncogene encodes a transcription factor which plays an important role in hematopoiesis. We detected truncated c-MYB mRNA (2.0 kb) and c-Myb protein (55 kDa) in the TK-6 cell line, which was established from a patient with chronic myelogenous leukemia in T cell blast crisis. Mutated c-MYB cDNA clone (WTK-1) was isolated from a TK-6 cell cDNA library and sequenced in its entirety. Compared with the wild-type human c-MYB sequence, the WTK-1 sequence diverged at the 3' ends of exons 9. A termination codon was present as the second codon downstream from the point of divergence and was followed by a previously unknown rearranged sequence. The conceptual protein encoded by WTK-1 (Myb(TK-6)) comprises 402 amino acids and lacks the negative regulatory domain of the normal c-Myb, reminiscent of the activated form of Myb protein. Luciferase reporter assay in NIH3T3 cells showed that the expression vector encoding Myb(TK-6) stimulated Myb-regulated mim-1 promoter more effectively than that encoding wild-type human c-Myb, suggesting that Myb(TK-6) is functional as a transcription factor, and thus as a potential transforming protein. Southern blot and mutant allele-specific polymerase chain reaction analyses showed that the same rearrangement of the c-MYB gene in TK-6 was present in late, but not in early, specimens obtained from the patient, indicating that this mutation had been acquired during disease progression.

Original languageEnglish
Pages (from-to)1422-1429
Number of pages8
JournalLeukemia
Volume12
Issue number9
DOIs
Publication statusPublished - 01-01-1998
Externally publishedYes

Fingerprint

Leukemia
Cell Line
Proto-Oncogene Proteins c-myb
Transcription Factors
Blast Crisis
Proteins
Proto-Oncogenes
Terminator Codon
Hematopoiesis
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Southern Blotting
Luciferases
Gene Library
Codon
Disease Progression
Exons
Complementary DNA
Clone Cells
Alleles
T-Lymphocytes

All Science Journal Classification (ASJC) codes

  • Hematology
  • Oncology
  • Cancer Research

Cite this

Tomita, A., Watanabe, T., Kosugi, H., Ohashi, H., Uchida, T., Kinoshita, T., ... Saito, H. (1998). Truncated c-Myb expression in the human leukemia cell line TK-6. Leukemia, 12(9), 1422-1429. https://doi.org/10.1038/sj.leu.2401113
Tomita, Akihiro ; Watanabe, T. ; Kosugi, H. ; Ohashi, H. ; Uchida, T. ; Kinoshita, T. ; Mizutani, S. ; Hotta, T. ; Murate, T. ; Seto, M. ; Saito, H. / Truncated c-Myb expression in the human leukemia cell line TK-6. In: Leukemia. 1998 ; Vol. 12, No. 9. pp. 1422-1429.
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Tomita, A, Watanabe, T, Kosugi, H, Ohashi, H, Uchida, T, Kinoshita, T, Mizutani, S, Hotta, T, Murate, T, Seto, M & Saito, H 1998, 'Truncated c-Myb expression in the human leukemia cell line TK-6', Leukemia, vol. 12, no. 9, pp. 1422-1429. https://doi.org/10.1038/sj.leu.2401113

Truncated c-Myb expression in the human leukemia cell line TK-6. / Tomita, Akihiro; Watanabe, T.; Kosugi, H.; Ohashi, H.; Uchida, T.; Kinoshita, T.; Mizutani, S.; Hotta, T.; Murate, T.; Seto, M.; Saito, H.

In: Leukemia, Vol. 12, No. 9, 01.01.1998, p. 1422-1429.

Research output: Contribution to journalArticle

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T1 - Truncated c-Myb expression in the human leukemia cell line TK-6

AU - Tomita, Akihiro

AU - Watanabe, T.

AU - Kosugi, H.

AU - Ohashi, H.

AU - Uchida, T.

AU - Kinoshita, T.

AU - Mizutani, S.

AU - Hotta, T.

AU - Murate, T.

AU - Seto, M.

AU - Saito, H.

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N2 - The c-MYB proto-oncogene encodes a transcription factor which plays an important role in hematopoiesis. We detected truncated c-MYB mRNA (2.0 kb) and c-Myb protein (55 kDa) in the TK-6 cell line, which was established from a patient with chronic myelogenous leukemia in T cell blast crisis. Mutated c-MYB cDNA clone (WTK-1) was isolated from a TK-6 cell cDNA library and sequenced in its entirety. Compared with the wild-type human c-MYB sequence, the WTK-1 sequence diverged at the 3' ends of exons 9. A termination codon was present as the second codon downstream from the point of divergence and was followed by a previously unknown rearranged sequence. The conceptual protein encoded by WTK-1 (Myb(TK-6)) comprises 402 amino acids and lacks the negative regulatory domain of the normal c-Myb, reminiscent of the activated form of Myb protein. Luciferase reporter assay in NIH3T3 cells showed that the expression vector encoding Myb(TK-6) stimulated Myb-regulated mim-1 promoter more effectively than that encoding wild-type human c-Myb, suggesting that Myb(TK-6) is functional as a transcription factor, and thus as a potential transforming protein. Southern blot and mutant allele-specific polymerase chain reaction analyses showed that the same rearrangement of the c-MYB gene in TK-6 was present in late, but not in early, specimens obtained from the patient, indicating that this mutation had been acquired during disease progression.

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Tomita A, Watanabe T, Kosugi H, Ohashi H, Uchida T, Kinoshita T et al. Truncated c-Myb expression in the human leukemia cell line TK-6. Leukemia. 1998 Jan 1;12(9):1422-1429. https://doi.org/10.1038/sj.leu.2401113