Multiple phosphorylation sites are important for RUNX1 activity in early hematopoiesis and T-cell differentiation

Mayumi Yoshimi, Susumu Goyama, Masahito Kawazu, Masahiro Nakagawa, Motoshi Ichikawa, Yoichi Imai, Keiki Kumano, Takashi Asai, James C. Mulloy, Andrew Kraft, Tsuyoshi Takahashi, Naoki Shirafuji, Mineo Kurokawa

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

RUNX1 is essential for definitive hematopoiesis and T-cell differentiation. It has been shown that RUNX1 is phosphorylated at specific serine and threonine residues by several kinase families. However, it remains unclear whether RUNX1 phosphorylation is absolutely required for its biological functions. Here, we evaluated hematopoietic activities of RUNX1 mutants with serine (S)/threonine (T) to alanine (A), aspartic acid (D), or glutamic acid (E) mutations at phosphorylation sites using primary culture systems. Consistent with the results of knockin mice, RUNX1-2A, carrying two phospho-deficient mutations at S276 and S293, retained hematopoietic activity. RUNX1-4A, carrying four mutations at S276, S293, T300, and S303, showed impaired T-cell differentiation activity, but retained the ability to rescue the defective early hematopoiesis of Runx1-deficient cells. Notably, RUNX1-5A, carrying five mutations at S276, S293, T300, S303, and S462, completely lost its hematopoietic activity. In contrast, the phospho-mimic proteins RUNX1-4D/E and RUNX1-5D/E exhibited normal function. Our study identifies multiple phosphorylation sites that are indispensable for RUNX1 activity in hematopoiesis.

Original languageEnglish (US)
Pages (from-to)1044-1050
Number of pages7
JournalEuropean Journal of Immunology
Volume42
Issue number4
DOIs
StatePublished - Apr 2012
Externally publishedYes

Fingerprint

Hematopoiesis
Cell Differentiation
Phosphorylation
T-Lymphocytes
Mutation
Threonine
Serine
D-Aspartic Acid
Alanine
Glutamic Acid
Phosphotransferases
Proteins

Keywords

  • Hematopoiesis
  • Phosphorylation
  • Posttranslational modification
  • RUNX1
  • T-cell differentiation

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy

Cite this

Yoshimi, M., Goyama, S., Kawazu, M., Nakagawa, M., Ichikawa, M., Imai, Y., ... Kurokawa, M. (2012). Multiple phosphorylation sites are important for RUNX1 activity in early hematopoiesis and T-cell differentiation. European Journal of Immunology, 42(4), 1044-1050. https://doi.org/10.1002/eji.201040746

Multiple phosphorylation sites are important for RUNX1 activity in early hematopoiesis and T-cell differentiation. / Yoshimi, Mayumi; Goyama, Susumu; Kawazu, Masahito; Nakagawa, Masahiro; Ichikawa, Motoshi; Imai, Yoichi; Kumano, Keiki; Asai, Takashi; Mulloy, James C.; Kraft, Andrew; Takahashi, Tsuyoshi; Shirafuji, Naoki; Kurokawa, Mineo.

In: European Journal of Immunology, Vol. 42, No. 4, 04.2012, p. 1044-1050.

Research output: Contribution to journalArticle

Yoshimi, M, Goyama, S, Kawazu, M, Nakagawa, M, Ichikawa, M, Imai, Y, Kumano, K, Asai, T, Mulloy, JC, Kraft, A, Takahashi, T, Shirafuji, N & Kurokawa, M 2012, 'Multiple phosphorylation sites are important for RUNX1 activity in early hematopoiesis and T-cell differentiation', European Journal of Immunology, vol. 42, no. 4, pp. 1044-1050. https://doi.org/10.1002/eji.201040746
Yoshimi, Mayumi ; Goyama, Susumu ; Kawazu, Masahito ; Nakagawa, Masahiro ; Ichikawa, Motoshi ; Imai, Yoichi ; Kumano, Keiki ; Asai, Takashi ; Mulloy, James C. ; Kraft, Andrew ; Takahashi, Tsuyoshi ; Shirafuji, Naoki ; Kurokawa, Mineo. / Multiple phosphorylation sites are important for RUNX1 activity in early hematopoiesis and T-cell differentiation. In: European Journal of Immunology. 2012 ; Vol. 42, No. 4. pp. 1044-1050.
@article{f0466e2b82b248d19a6ef86295c45b7c,
title = "Multiple phosphorylation sites are important for RUNX1 activity in early hematopoiesis and T-cell differentiation",
abstract = "RUNX1 is essential for definitive hematopoiesis and T-cell differentiation. It has been shown that RUNX1 is phosphorylated at specific serine and threonine residues by several kinase families. However, it remains unclear whether RUNX1 phosphorylation is absolutely required for its biological functions. Here, we evaluated hematopoietic activities of RUNX1 mutants with serine (S)/threonine (T) to alanine (A), aspartic acid (D), or glutamic acid (E) mutations at phosphorylation sites using primary culture systems. Consistent with the results of knockin mice, RUNX1-2A, carrying two phospho-deficient mutations at S276 and S293, retained hematopoietic activity. RUNX1-4A, carrying four mutations at S276, S293, T300, and S303, showed impaired T-cell differentiation activity, but retained the ability to rescue the defective early hematopoiesis of Runx1-deficient cells. Notably, RUNX1-5A, carrying five mutations at S276, S293, T300, S303, and S462, completely lost its hematopoietic activity. In contrast, the phospho-mimic proteins RUNX1-4D/E and RUNX1-5D/E exhibited normal function. Our study identifies multiple phosphorylation sites that are indispensable for RUNX1 activity in hematopoiesis.",
keywords = "Hematopoiesis, Phosphorylation, Posttranslational modification, RUNX1, T-cell differentiation",
author = "Mayumi Yoshimi and Susumu Goyama and Masahito Kawazu and Masahiro Nakagawa and Motoshi Ichikawa and Yoichi Imai and Keiki Kumano and Takashi Asai and Mulloy, {James C.} and Andrew Kraft and Tsuyoshi Takahashi and Naoki Shirafuji and Mineo Kurokawa",
year = "2012",
month = "4",
doi = "10.1002/eji.201040746",
language = "English (US)",
volume = "42",
pages = "1044--1050",
journal = "European Journal of Immunology",
issn = "0014-2980",
publisher = "Wiley-VCH Verlag",
number = "4",

}

TY - JOUR

T1 - Multiple phosphorylation sites are important for RUNX1 activity in early hematopoiesis and T-cell differentiation

AU - Yoshimi, Mayumi

AU - Goyama, Susumu

AU - Kawazu, Masahito

AU - Nakagawa, Masahiro

AU - Ichikawa, Motoshi

AU - Imai, Yoichi

AU - Kumano, Keiki

AU - Asai, Takashi

AU - Mulloy, James C.

AU - Kraft, Andrew

AU - Takahashi, Tsuyoshi

AU - Shirafuji, Naoki

AU - Kurokawa, Mineo

PY - 2012/4

Y1 - 2012/4

N2 - RUNX1 is essential for definitive hematopoiesis and T-cell differentiation. It has been shown that RUNX1 is phosphorylated at specific serine and threonine residues by several kinase families. However, it remains unclear whether RUNX1 phosphorylation is absolutely required for its biological functions. Here, we evaluated hematopoietic activities of RUNX1 mutants with serine (S)/threonine (T) to alanine (A), aspartic acid (D), or glutamic acid (E) mutations at phosphorylation sites using primary culture systems. Consistent with the results of knockin mice, RUNX1-2A, carrying two phospho-deficient mutations at S276 and S293, retained hematopoietic activity. RUNX1-4A, carrying four mutations at S276, S293, T300, and S303, showed impaired T-cell differentiation activity, but retained the ability to rescue the defective early hematopoiesis of Runx1-deficient cells. Notably, RUNX1-5A, carrying five mutations at S276, S293, T300, S303, and S462, completely lost its hematopoietic activity. In contrast, the phospho-mimic proteins RUNX1-4D/E and RUNX1-5D/E exhibited normal function. Our study identifies multiple phosphorylation sites that are indispensable for RUNX1 activity in hematopoiesis.

AB - RUNX1 is essential for definitive hematopoiesis and T-cell differentiation. It has been shown that RUNX1 is phosphorylated at specific serine and threonine residues by several kinase families. However, it remains unclear whether RUNX1 phosphorylation is absolutely required for its biological functions. Here, we evaluated hematopoietic activities of RUNX1 mutants with serine (S)/threonine (T) to alanine (A), aspartic acid (D), or glutamic acid (E) mutations at phosphorylation sites using primary culture systems. Consistent with the results of knockin mice, RUNX1-2A, carrying two phospho-deficient mutations at S276 and S293, retained hematopoietic activity. RUNX1-4A, carrying four mutations at S276, S293, T300, and S303, showed impaired T-cell differentiation activity, but retained the ability to rescue the defective early hematopoiesis of Runx1-deficient cells. Notably, RUNX1-5A, carrying five mutations at S276, S293, T300, S303, and S462, completely lost its hematopoietic activity. In contrast, the phospho-mimic proteins RUNX1-4D/E and RUNX1-5D/E exhibited normal function. Our study identifies multiple phosphorylation sites that are indispensable for RUNX1 activity in hematopoiesis.

KW - Hematopoiesis

KW - Phosphorylation

KW - Posttranslational modification

KW - RUNX1

KW - T-cell differentiation

UR - http://www.scopus.com/inward/record.url?scp=84860248542&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84860248542&partnerID=8YFLogxK

U2 - 10.1002/eji.201040746

DO - 10.1002/eji.201040746

M3 - Article

VL - 42

SP - 1044

EP - 1050

JO - European Journal of Immunology

JF - European Journal of Immunology

SN - 0014-2980

IS - 4

ER -