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DOCK2 associates with CrkL and regulates Rac1 in human leukemia cell lines

Nishihara, Hiroshi, Maeda, Masae, Oda, Atsushi, Tsuda, Masumi, Sawa, Hirofumi, Nagashima, Kazuo, Tanaka, Shinya·2002·Blood
DOI 10.1182/blood-2001-11-0032
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Abstract

The CDM (ced-5 of Caenorhabditis elegans,DOCK180 [downstream of Crkwith molecular weight of 180 kDa] of humans, andmyoblast city of Drosophila melanogaster) family of proteins has been shown to play a pivotal role in the integrin-mediated signaling pathway under the regulation of an adaptor moleculec-CT10–related kinase II (c–Crk-II) in adherent cells. Recently, hematopoietic cell–specific CDM protein DOCK2 has been shown to be indispensable for lymphocyte migration. However, the regulatory mechanism for DOCK2 is still unknown because DOCK2 lacks a c–Crk-II binding consensus motif. In this study, we demonstrated that DOCK2 bound to CrkL, which is present exclusively in hematopoietic cells both in vivo and in vitro, and we also found that 2 separate regions of DOCK2 contributed to its binding to Src homology 3 (SH3) domain of CrkL. Colocalization of DOCK2 with Crk-like (CrkL) and F-actin was shown by immunocytochemical analysis with the use of Jurkat cells. We also found that CrkL-induced activation of small guanine triphosphatase (GTPase) Rac1 was significantly inhibited by the DOCK2-dCS mutant in 293T cells. Furthermore, the association of DOCK2 and Vav, the guanine-nucleotide exchanging factor (GEF) for Rac1, was demonstrated in Jurkat cells. Finally, the stable expression of DOCK2-dCS mutant in Jurkat cells was shown to reduce cell attachment. These data suggest the presence of a novel protein complex of CrkL, DOCK2, and Vav to regulate Rac1 in leukemia cell lines.

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Research works printed but not linked: 37

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Metadata

Authors
Nishihara, H., Maeda, M., Oda, A., Tsuda, M., Sawa, H., Nagashima, K., & Tanaka, S.
Affiliations
From the Laboratory of Molecular and Cellular Pathology, and the Laboratory of Environmental Biology, Hokkaido University School of Medicine, Sapporo, Japan; and CREST, Japan Science and Technology Corporation.
Venue
Blood
Publisher
American Society of Hematology
Published
2002-12-01
Volume
100
Issue
12
Pages
3968-3974
ISSN
  • 1528-0020
  • 0006-4971
DOI
10.1182/blood-2001-11-0032
Cited by
64
References
1
Language
en

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