PMID- 11090054
OWN - NLM
STAT- MEDLINE
DCOM- 20010201
LR  - 20210216
IS  - 0006-4971 (Print)
IS  - 0006-4971 (Linking)
VI  - 96
IP  - 12
DP  - 2000 Dec 1
TI  - Differential stimulation of c-Kit mutants by membrane-bound and soluble Steel 
      Factor correlates with leukemic potential.
PG  - 3734-42
AB  - The authors investigated the roles of PI3-kinase and PLC-gamma in stimulation by 
      Steel Factor (SLF) through c-Kit. c-Kit mutants YF719, YF728, and a YF719/YF728 
      double mutant were expressed in 32D myelomonocytic cells. KitYF719 fails to 
      recruit PI3-kinase after stimulation with SLF, whereas KitYF728 fails to 
      stimulate PLC-gamma phosphorylation or mobilize Ca(++). Both single mutants 
      responded mitogenically to soluble SLF (sSLF) in a manner indistinguishable from 
      wild type (WT), although sSLF failed to stimulate or promote the survival of 
      cells expressing the double mutant. In contrast, although cells expressing WT or 
      YF719 were mitogenically stimulated by membrane-bound SLF (mSLF), stimulation of 
      cells expressing KitYF728 was impaired. Similarly, cells expressing WT or YF719 
      receptors were stimulated by plate-bound anti-Kit antibodies, whereas cells 
      expressing the YF728 receptor were not stimulated. Neomycin sulfate, a PLC 
      antagonist, inhibited cells expressing YF719 receptors stimulated by sSLF. 
      Neomycin also inhibited cells expressing the WT receptor that were stimulated by 
      mSLF or immobilized anti-Kit antibodies but did not inhibit stimulation of cells 
      expressing WT or YF719 receptors by sSLF. 32D cells expressing KitWT, KitYF719, 
      or KitYF728 were injected into mice and the presence of cells was evaluated by 
      colony assays 6 to 7 weeks later. Although both KitWT and KitYF719 expressing 
      cells could be recovered from the spleen and bone marrow, recovery of KitYF728 
      cells from these organs was severely reduced. These results indicate that Kit 
      tyrosine 728 is of particular importance for mitogenic stimulation by mSLF or 
      immobilized ligand and is required for full maintenance of cells in vivo, likely 
      through activation of PLC-gamma. (Blood. 2000;96:3734-3742)
FAU - Gommerman, J L
AU  - Gommerman JL
AD  - The Arthritis and Immune Disorder Research Centre, The Toronto Hospital, 
      University of Toronto, Toronto, Ontario, Canada.
FAU - Sittaro, D
AU  - Sittaro D
FAU - Klebasz, N Z
AU  - Klebasz NZ
FAU - Williams, D A
AU  - Williams DA
FAU - Berger, S A
AU  - Berger SA
LA  - eng
GR  - 2RO1 DK 48605/DK/NIDDK NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Blood
JT  - Blood
JID - 7603509
RN  - 0 (Antibodies)
RN  - 0 (Isoenzymes)
RN  - 0 (Membrane Proteins)
RN  - 0 (Mitogens)
RN  - 0 (Protein Subunits)
RN  - 0 (Stem Cell Factor)
RN  - EC 2.7.1.- (Phosphatidylinositol 3-Kinases)
RN  - EC 2.7.10.1 (Proto-Oncogene Proteins c-kit)
RN  - EC 3.1.4.- (Type C Phospholipases)
RN  - EC 3.1.4.3 (Phospholipase C gamma)
RN  - I16QD7X297 (Neomycin)
SB  - IM
MH  - Animals
MH  - Antibodies/metabolism
MH  - Calcium Signaling/drug effects
MH  - Enzyme Activation/physiology
MH  - Humans
MH  - Isoenzymes/chemistry/metabolism/physiology
MH  - Leukemia/enzymology/*etiology
MH  - Membrane Proteins/*pharmacology
MH  - Mice
MH  - Mitogens/pharmacology
MH  - Mutagenesis, Site-Directed
MH  - Neomycin/pharmacology
MH  - Phosphatidylinositol 3-Kinases/chemistry/metabolism/physiology
MH  - Phospholipase C gamma
MH  - Phosphorylation
MH  - Protein Subunits
MH  - Proto-Oncogene Proteins c-kit/*drug effects/genetics/immunology/metabolism
MH  - Signal Transduction/drug effects
MH  - Solubility
MH  - Stem Cell Factor/*pharmacology
MH  - Transfection
MH  - Tumor Cells, Cultured/drug effects
MH  - Type C Phospholipases/chemistry/metabolism/physiology
MH  - src Homology Domains/physiology
EDAT- 2000/11/23 11:00
MHDA- 2001/02/28 10:01
CRDT- 2000/11/23 11:00
PHST- 2000/11/23 11:00 [pubmed]
PHST- 2001/02/28 10:01 [medline]
PHST- 2000/11/23 11:00 [entrez]
AID - S0006-4971(20)48212-6 [pii]
PST - ppublish
SO  - Blood. 2000 Dec 1;96(12):3734-42.