PMID- 10477722
OWN - NLM
STAT- MEDLINE
DCOM- 19991012
LR  - 20061115
IS  - 0006-4971 (Print)
IS  - 0006-4971 (Linking)
VI  - 94
IP  - 6
DP  - 1999 Sep 15
TI  - Identification of a 14-3-3 binding sequence in the common beta chain of the
      granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), 
      and IL-5 receptors that is serine-phosphorylated by GM-CSF.
PG  - 1933-42
AB  - The common beta chain (beta(c)) of the granulocyte-macrophage colony-stimulating 
      factor (GM-CSF), interleukin-3 (IL-3), and IL-5 receptors is the major signaling 
      subunit of these receptors coupling ligand binding to multiple biological
      activities. It is thought that these multiple functions arise as a consequence of
      the recruitment of specific signaling molecules to tyrosine-phosphorylated
      residues in the cytoplasmic domain of beta(c). However, the contribution of
      serine phosphorylation in beta(c) to the recruitment of signaling molecules is
      not known. We show here the identification of a phosphoserine motif in the
      cytoplasmic domain of beta(c) that interacts with the adaptor protein 14-3-3zeta.
      Coimmunoprecipitation and pull-down experiments with a glutathione S-transferase 
      (GST):14-3-3zeta fusion protein showed that 14-3-3 directly associates with
      beta(c) but not the GM-CSF receptor alpha chain. C-terminal truncation mutants of
      beta(c) further showed that a region between amino acids 544 and 626 in beta(c)
      was required for its association with 14-3-3zeta. This region contains the
      sequence (582)HSRSLP(587), which closely resembles the RSXSXP (where S is
      phosphorylated) consensus 14-3-3 binding site identified in a number of signaling
      molecules, including Raf-1. Significantly, substitution of (582)HSRSLP(587) for
      EFAAAA completely abolished interaction of beta(c) with GST-14-3-3zeta.
      Furthermore, the interaction of beta(c) with GST-14-3-3 was greatly reduced in
      the presence of a peptide containing the 14-3-3 binding site, but only when
      (585)Ser was phosphorylated. Direct binding experiments showed that the peptide
      containing phosphorylated (585)Ser bound 14-3-3zeta with an affinity of 150
      nmol/L. To study the regulation of (585)S phosphorylation in vivo, we raised
      antibodies that specifically recognized (585)Ser-phosphorylated beta(c). Using
      these antibodies, we showed that GM-CSF stimulation strongly upregulated (585)Ser
      phosphorylation in M1 myeloid leukemic cells. The proximity of the SHC-binding
      site ((577)Tyr) to the 14-3-3-binding site ((582)HSRSLP(587)) and their
      conservation between mouse, rat, and human beta(c) but not in other cytokine
      receptors suggest that they form a distinct motif that may subserve specialized
      functions associated with the GM-CSF, IL-3, and IL-5 receptors.
FAU - Stomski, F C
AU  - Stomski FC
AD  - The Cytokine Receptor Laboratory, The Hanson Centre for Cancer Research and
      Institute of Medical and Veterinary Science, Adelaide, Australia.
FAU - Dottore, M
AU  - Dottore M
FAU - Winnall, W
AU  - Winnall W
FAU - Guthridge, M A
AU  - Guthridge MA
FAU - Woodcock, J
AU  - Woodcock J
FAU - Bagley, C J
AU  - Bagley CJ
FAU - Thomas, D T
AU  - Thomas DT
FAU - Andrews, R K
AU  - Andrews RK
FAU - Berndt, M C
AU  - Berndt MC
FAU - Lopez, A F
AU  - Lopez AF
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Blood
JT  - Blood
JID - 7603509
RN  - 0 (14-3-3 Proteins)
RN  - 0 (Macromolecular Substances)
RN  - 0 (Proteins)
RN  - 0 (Receptors, Granulocyte-Macrophage Colony-Stimulating Factor)
RN  - 0 (Receptors, Interleukin)
RN  - 0 (Receptors, Interleukin-3)
RN  - 0 (Receptors, Interleukin-5)
RN  - 0 (Recombinant Proteins)
RN  - 17885-08-4 (Phosphoserine)
RN  - 83869-56-1 (Granulocyte-Macrophage Colony-Stimulating Factor)
RN  - EC 1.14.16.2 (Tyrosine 3-Monooxygenase)
SB  - AIM
SB  - IM
MH  - 14-3-3 Proteins
MH  - Amino Acid Sequence
MH  - Amino Acid Substitution
MH  - Animals
MH  - Binding Sites
MH  - Granulocyte-Macrophage Colony-Stimulating Factor/*metabolism
MH  - Humans
MH  - Kinetics
MH  - Macromolecular Substances
MH  - Mice
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Phosphorylation
MH  - Phosphoserine/*metabolism
MH  - Point Mutation
MH  - Proteins/*chemistry/*metabolism
MH  - Rats
MH  - Receptors, Granulocyte-Macrophage Colony-Stimulating Factor/chemistry/*metabolism
MH  - Receptors, Interleukin/chemistry/*metabolism
MH  - Receptors, Interleukin-3/chemistry/*metabolism
MH  - Receptors, Interleukin-5
MH  - Recombinant Proteins/chemistry/metabolism
MH  - Sequence Alignment
MH  - Sequence Homology, Amino Acid
MH  - *Tyrosine 3-Monooxygenase
EDAT- 1999/09/09 00:00
MHDA- 1999/09/09 00:01
CRDT- 1999/09/09 00:00
PHST- 1999/09/09 00:00 [pubmed]
PHST- 1999/09/09 00:01 [medline]
PHST- 1999/09/09 00:00 [entrez]
PST - ppublish
SO  - Blood. 1999 Sep 15;94(6):1933-42.