|
|
 Previous Article | Table of Contents | Next Article 
Blood, 15 June 2001, Vol. 97, No. 12, pp. 3768-3775
HEMATOPOIESIS
The adapter protein CrkL associates with CD34
Donna M. Felschow,
Megan L. McVeigh,
Gerard T. Hoehn,
Curt I. Civin, and
Mary Jo Fackler
From the Johns Hopkins Oncology Center, Johns Hopkins
University School of Medicine, Bunting-Blaustein Cancer Research
Building, Baltimore, MD.
CD34 is a cell-surface transmembrane protein expressed specifically
at the stem/progenitor stage of lymphohematopoietic development that
appears to regulate adhesion. To elucidate intracellular signals
modified by CD34, we designed and constructed
glutathione-S-transferase (GST)- fusion proteins of the
intracellular domain of full-length CD34 (GST-CD34ifull).
Precipitation of cell lysates using GST-CD34ifull identified proteins of molecular mass 39, 36, and 33 kd that
constitutively associated with CD34 and a 45-kd protein that associated
with CD34 after adhesion. By Western analysis, we identified the 39-kd protein as CrkL. In vivo, CrkL was coimmunoprecipitated with CD34 using
CD34 antibodies, confirming the association between CrkL and CD34. CD34
peptide inhibition assays demonstrated that CrkL interacts at a
membrane-proximal region of the CD34 tail. To identify the CrkL domain
responsible for interaction with CD34, we generated GST-fusion
constructs of adapter proteins including GST-CrkL3' (C-terminal
SH3) and GST-CrkL5' (N-terminal SH2SH3). Of these fusion proteins, only GST-CrkL3' could precipitate endogenously expressed CD34, suggesting that CD34 binds the C-terminal SH3 domain of
CrkL. Interestingly, there appears to be differential specificity
between CrkL and CrkII for CD34, because GST-CD34ifull did
not precipitate CrkII, a highly homologous Crk family member. Furthermore, GST-CD34ifull did not bind c-Abl, c-Cbl, C3G,
or paxillin proteins that are known to associate with CrkL, suggesting that CD34 directly interacts with the CrkL protein.
CD34ifull association with Grb or Shc adapter proteins was
not detected. Our investigations shed new light on signaling pathways
of CD34 by demonstrating that CD34 couples to the hematopoietic adapter protein CrkL.
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
J. S. Nielsen and K. M. McNagny
Novel functions of the CD34 family
J. Cell Sci.,
November 15, 2008;
121(22):
3683 - 3692.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. M. Acevedo, I. Londono, M. Oubaha, L. Ghitescu, and M. Bendayan
Glomerular CD34 Expression in Short- and Long-term Diabetes
J. Histochem. Cytochem.,
June 1, 2008;
56(6):
605 - 614.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. S. Trempus, R. J. Morris, M. Ehinger, A. Elmore, C. D. Bortner, M. Ito, G. Cotsarelis, J. G.W. Nijhof, J. Peckham, N. Flagler, et al.
CD34 Expression by Hair Follicle Stem Cells Is Required for Skin Tumor Development in Mice
Cancer Res.,
May 1, 2007;
67(9):
4173 - 4181.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. C. Tan, S. G.B. Furness, H. Merkens, S. Lin, M. L. McCoy, C. D. Roskelley, J. Kast, and K. M. McNagny
Na+/H+ Exchanger Regulatory Factor-1 Is a Hematopoietic Ligand for a Subset of the CD34 Family of Stem Cell Surface Proteins
Stem Cells,
May 1, 2006;
24(5):
1150 - 1161.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. C. Dooley, B. K. Oppenlander, and M. Xiao
Analysis of Primitive CD34- and CD34+ Hematopoietic Cells from Adults: Gain and Loss of CD34 Antigen by Undifferentiated Cells Are Closely Linked to Proliferative Status in Culture
Stem Cells,
July 1, 2004;
22(4):
556 - 569.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Nishihara, M. Maeda, A. Oda, M. Tsuda, H. Sawa, K. Nagashima, and S. Tanaka
DOCK2 associates with CrkL and regulates Rac1 in human leukemia cell lines
Blood,
December 1, 2002;
100(12):
3968 - 3974.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
