This project aims to investigate the function of the receptors for granulocyte-macrophage colony stimulating factor (GM-CSF), Interleukin-3 (IL-3) and Interleukin-5 (IL-5). These factors exert strong proliferative and differentiative effects on myeloid hemopoietic cells, and moreover, autocrine GM-CSF production has been implicated in some leukemias, while IL-5 is involved in many allergic reactions. The cell-surface receptors for GM-CSF, IL-3 and IL-5 are members of the cytokine receptor family and are comprised of two different subunits, alpha and beta. The alpha subunit is unique to each factor while the beta subunit (h-beta-c) is shared by the three factors. Specifically, it is proposed to utilize a unique set of constitutively active mutants of the h-beta-c to examine the structure, composition and activity of the functional receptor complexes. These studies will test a model which predicts that there are actually two distinct types of functional receptor complex. This will be carried out by: (i) Examining association between receptor subunits expressed in cell lines; (ii) Studying the molecular structure of a critical region of h-beta-c in both normal and mutant forms. This will use both directed mutations and nuclear magnetic resonance spectroscopy; (iii) Attempting to correlate different receptor subunit associations with tyrosine phosphorylation of h-beta-c. In addition, it is proposed that novel reagents (e.g., monoclonal antibodies and peptides) can be generated which interact with critical regions of h-beta-c and which can trigger its signalling activity. As well as providing tools to probe receptor structure, these reagents may provide a proof-of-principle for the isolation of novel agonists for these and other members of the cytokine receptor family.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL060657-03
Application #
6184541
Study Section
Hematology Subcommittee 2 (HEM)
Project Start
1998-08-01
Project End
2001-07-31
Budget Start
2000-08-01
Budget End
2001-07-31
Support Year
3
Fiscal Year
2000
Total Cost
$127,964
Indirect Cost
Name
Institute of Medical and Vet Science
Department
Type
DUNS #
740198254
City
Adelaide
State
Country
Australia
Zip Code
5000
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Perugini, Michelle; Brown, Anna L; Salerno, Diana G et al. (2010) Alternative modes of GM-CSF receptor activation revealed using activated mutants of the common beta-subunit. Blood 115:3346-53
Perugini, M; Kok, C H; Brown, A L et al. (2009) Repression of Gadd45alpha by activated FLT3 and GM-CSF receptor mutants contributes to growth, survival and blocked differentiation. Leukemia 23:729-38
Brown, Anna L; Wilkinson, Christopher R; Waterman, Scott R et al. (2006) Genetic regulators of myelopoiesis and leukemic signaling identified by gene profiling and linear modeling. J Leukoc Biol 80:433-47
D'Andrea, Richard J; Sadlon, Timothy J; Gonda, Thomas J (2004) Overlapping motifs in the membrane-proximal region of cytokine receptor accessory and signaling subunits. Cytokine Growth Factor Rev 15:83-5
McClure, B J; Woodcock, J M; Harrison-Findik, D et al. (2001) GM-CSF binding to its receptor induces oligomerisation of the common beta-subunit. Cytokine 13:240-3
Butcher, C; D'Andrea, R J (2000) Molecular aspects of polycythemia vera (review). Int J Mol Med 6:243-52
Mulhern, T D; Lopez, A F; D'Andrea, R J et al. (2000) The solution structure of the cytokine-binding domain of the common beta-chain of the receptors for granulocyte-macrophage colony-stimulating factor, interleukin-3 and interleukin-5. J Mol Biol 297:989-1001
D'Andrea, R J; Gonda, T J (2000) A model for assembly and activation of the GM-CSF, IL-3 and IL-5 receptors: insights from activated mutants of the common beta subunit. Exp Hematol 28:231-43