PMID- 10608804
OWN - NLM
STAT- MEDLINE
DCOM- 20000208
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 53
DP  - 1999 Dec 31
TI  - The adapter protein Crkl links Cbl to C3G after integrin ligation and enhances
      cell migration.
PG  - 37525-32
AB  - Crkl, an SH2-SH3-SH3 adapter protein, is one of the major tyrosine
      phosphoproteins detected in cells from patients with chronic myelogenous
      leukemia. Crkl binds to BCR/ABL through its N-terminal SH3 domain and is known to
      interact with several signaling proteins that have been implicated in integrin
      signaling, including Cbl, Cas, Hef-1, and paxillin. We have previously shown that
      overexpression of Crkl enhances adhesion to extracellular matrix proteins through
      beta(1) integrins. In this study, the effects of Crkl on spontaneous and
      chemokine-directed migration of the hematopoietic cell line Ba/F3 were examined. 
      Full-length, SH2-, and SH3(N)-domain deletion mutants of Crkl were expressed
      transiently as fusion proteins with green fluorescent protein. Successfully
      transfected cells were isolated by fluorescence-activated cell sorting. The
      ability of these cells to migrate across a fibronectin-coated membrane, either
      spontaneously or in response to the chemokine stromal-derived factor-1alpha, was 
      determined. Cells expressing green fluorescent protein alone were not
      distinguishable from untransfected or mock transfected Ba/F3 cells. However,
      Ba/F3 cells overexpressing full-length Crkl were found to have an increase in
      spontaneous migration of 2.8 +/- 0.6-fold in seven independent assays. The
      enhancement of migration required both the SH2 domain and the N-terminal SH3
      domain. Migration in response to stromal-derived factor-1alpha was not
      significantly enhanced by overexpression of Crkl. Overexpression of Crkii also
      augmented spontaneous migration but to a lesser degree than did Crkl. Because the
      SH2 domain was required for enhanced migration, we looked for changes in
      phosphotyrosine containing proteins coprecipitating with Crkl, but not Crkl
      DeltaSH2, after integrin cross-linking. Full-length Crkl, but not CrklDeltaSH2,
      coprecipitated with a single major tyrosine phosphoprotein with an M(r) of
      approximately 120 kDa, identified as Cbl. The major Crkl SH3-binding protein in
      these cells was found to be the guanine nucleotide exchange factor, C3G.
      Interestingly, overexpression of C3G also enhanced migration, suggesting that a
      Cbl-Crkl-C3G complex may be involved in migration signaling in Ba/F3 cells. These
      data suggest that Crkl is involved in signaling pathways that regulate migration,
      possibly through a complex with Cbl and C3G.
FAU - Uemura, N
AU  - Uemura N
AD  - Department of Adult Oncology, Dana-Farber Cancer Institute, Brigham and Women's
      Hospital and Harvard Medical School, Boston, Massachusetts 02115, USA.
FAU - Griffin, J D
AU  - Griffin JD
LA  - eng
GR  - CA36167/CA/NCI NIH HHS/United States
GR  - DK560654/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Guanine Nucleotide-Releasing Factor 2)
RN  - 0 (Integrin beta1)
RN  - 0 (Proto-Oncogene Proteins)
RN  - 0 (Proto-Oncogene Proteins c-crk)
RN  - EC 2.3.2.27 (Proto-Oncogene Proteins c-cbl)
RN  - EC 2.3.2.27 (Ubiquitin-Protein Ligases)
RN  - EC 2.7.- (Protein Kinases)
RN  - EC 6.3.2.- (CBL protein, human)
RN  - EC 6.3.2.- (Cbl protein, mouse)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Cell Movement/*physiology
MH  - Guanine Nucleotide-Releasing Factor 2/*metabolism
MH  - Humans
MH  - Integrin beta1/*metabolism
MH  - Mice
MH  - Protein Binding
MH  - Protein Kinases/*metabolism/physiology
MH  - Proto-Oncogene Proteins/*metabolism
MH  - Proto-Oncogene Proteins c-cbl
MH  - Proto-Oncogene Proteins c-crk
MH  - *Ubiquitin-Protein Ligases
EDAT- 1999/12/23 00:00
MHDA- 1999/12/23 00:01
CRDT- 1999/12/23 00:00
PHST- 1999/12/23 00:00 [pubmed]
PHST- 1999/12/23 00:01 [medline]
PHST- 1999/12/23 00:00 [entrez]
AID - 10.1074/jbc.274.53.37525 [doi]
AID - S0021-9258(19)52924-6 [pii]
PST - ppublish
SO  - J Biol Chem. 1999 Dec 31;274(53):37525-32. doi: 10.1074/jbc.274.53.37525.