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Originally published In Press as doi:10.1074/jbc.M101488200 on September 10, 2001
J. Biol. Chem., Vol. 276, Issue 45, 41803-41809, November 9, 2001
Cell Cycle-dependent Interaction of Mad2 with
Conserved Box1/2 Region of Human Granulocyte-Macrophage
Colony-stimulating Factor Receptor Common c*
Mitsuo
Takeda §,
Naoshi
Dohmae¶,
Koji
Takio¶,
Ken-ichi
Arai §, and
Sumiko
Watanabe
From the Department of Molecular and Developmental
Biology, Institute of Medical Science, § Core Research for
Evolutional Science and Technology, Japan Science and Technology
Corporation, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo
108-8639, Japan, and the ¶ Biomolecular Characterization Division,
RIKEN (Institute of Physical and Chemical Research), 2-1, Hirosawa,
Wako-shi, Saitama 351-0198, Japan
Box1 and 2 (box1/2) are conserved cytoplasmic
motifs located in the membrane proximal region of cytokine receptors,
including the human granulocyte-macrophage colony-stimulating
factor (GM-CSF) receptor common c. Deletion of box1/2 abrogated all
the examined activities of GM-CSF, and this phenomenon is explained by
the loss of binding by Jak2. To test if a molecule other than Jak2 interacting with the box1/2 region plays a role in GM-CSF receptor signal transduction, we screened for molecules interacting with the
box1/2 region by a pull-down assay using recombinant purified protein
of GST fused with the c box1/2 region and a Ba/F3 cell lysate. The
mouse homologue of Mad2 protein, which plays an important role in the M
phase of the cell cycle, was revealed to associate with the box1/2
region specifically. Peptides corresponding to the box1
sequence also bound to Mad2, and mutation of the box1 decreased the Mad2 interaction. Deletion analysis indicated that interaction with box1/2 occurred through the C-terminal portion of
Mad2. Mad2 is known to change affinity for binding partners cell cycle
dependently. Binding affinity of Mad2 to box1/2 increased in the late M
phase, suggesting the possibility that GM-CSF participates in
regulation of the M phase check point through interaction with Mad2.
*
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of
Molecular and Developmental Biology, Inst. of Medical Science,
University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan. Tel.: 81-3-5449-5660; Fax: 81-3-5449-5424; E-mail:
sumiko@ims.u-tokyo.ac.jp.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.

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