Determining c-Myb protein levels can isolate functional hematopoietic stem cell subtypes

Hiroshi Sakamoto, Naoki Takeda, Fumio Arai, Kentaro Hosokawa, Paloma Garcia, Toshio Suda, Jon Frampton, Minetaro Ogawa

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

The transcription factor c-Myb was originally identified as a transforming oncoprotein encoded by two avian leukemia viruses. Subsequently, through the generation of mouse models that affect its expression, c-Myb has been shown to be a key regulator of hematopoiesis, including having critical roles in hematopoietic stem cells (HSCs). The precise function of c-Myb in HSCs although remains unclear. We have generated a novel c-myb allele in mice that allows direct observation of c-Myb protein levels in single cells. Using this reporter line we demonstrate that subtypes of HSCs can be isolated based upon their respective c-Myb protein expression levels. HSCs expressing low levels of c-Myb protein (c-MyblowHSC) appear to represent the most immature, dormant HSCs and they are a predominant component of HSCs that retain bromodeoxyuridine labeling. Hematopoietic stress, induced by 5-fluorouracil ablation, revealed that in this circumstance c-Myb-expressing cells become critical for multilineage repopulation. The discrimination of HSC subpopulations based on c-Myb protein levels is not reflected in the levels of c-myb mRNA, there being no more than a 1.3-fold difference comparing c-Myblow and c-MybhighHSCs. This illustrates how essential it is to include protein studies when aiming to understand the regulatory networks that control stem cell behavior.

Original languageEnglish
Pages (from-to)479-490
Number of pages12
JournalStem Cells
Volume33
Issue number2
DOIs
Publication statusPublished - 2015 Feb 1

Keywords

  • Animals
  • Cell proliferation
  • Genetically modified
  • Hematopoiesis
  • Hematopoietic stem cell transplantation
  • Hematopoietic stem cells
  • Proto-oncogene proteins c-myb

ASJC Scopus subject areas

  • Molecular Medicine
  • Developmental Biology
  • Cell Biology

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  • Cite this

    Sakamoto, H., Takeda, N., Arai, F., Hosokawa, K., Garcia, P., Suda, T., Frampton, J., & Ogawa, M. (2015). Determining c-Myb protein levels can isolate functional hematopoietic stem cell subtypes. Stem Cells, 33(2), 479-490. https://doi.org/10.1002/stem.1855