The innate immune response is an important line of defense against pathogenic agents and is also an integral component of the wound repair process. In vivo, the cytokines, tumor necrosis factor alpha (TNFalpha) and interleukin (IL 1) activate the innate immune response by increasing NF-kappaB dependent expression of genes. For example, interleukin-8 recruits circulating phagocytic cells and E-selectin facilitates adherence of recruited phagocytes. Together changes in IL-8 and E-selectin gene expression are key events that occur during the innate immune response. A high degree of sequence similarity is shared between the mammalian NF- kappaB/c-rel family and its cytoplasmic inhibitor IkappaBalpha, and the Drosophila Dorsal protein and its regulator Cactus. Genetic analyses of Dorsal have defined components of a signaling pathway for Dorsal activation, including a serine/threonine protein kinases, Pelle, upstream of Dorsal and Cactus. We demonstrated that this pathway is conserved in mammals through the isolation of a cDNA that encodes a novel mouse protein highly related to Pelle, mPLK (mouse Pelle-like protein kinase). Over- expression of the mPLK cDNA in fibroblasts leads to transactivation of the NF-kappaB dependent IL-8 and E-selectin gene promoters. Catalytically inactive mPLK functions as a dominant-negative and blocks TNFalpha mediated transactivation of the IL-8 gene promoter. Moreover in unstimulated cells, mPLK is complexed with the type I tumor necrosis factor receptor (TNFRI) which is required for TNFalpha induced NF kappaB activity. Current TNFRI signaling models predict that TNFRI mediated induction of NF-kappaB activity is mediated by the TNF receptor associated death domain protein (TRADD), which associates with TNFRI after TNFalpha stimulation. However our recent data, demonstrate that mPLK is also a mediator through which TNFRI can induce NF-kappaB activity. The studies in this proposal are aimed at testing the hypothesis that mPLK is a TRADD- independent pathway through which TNFRI can induce NF-kappaB activity. This will be accomplished by: (1) Identifying TNFRI-mPLK complex components; (2) Determining if mPLK signaling activity is TRADD dependent; (3) Identifying mPLK-NIK (NF-kappaB inducing kinase) complex components; (4) Determining if TNFRI and mPLK activities are interdependent; and (5) Identifying additional mPLK regulators.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI042798-03
Application #
6341702
Study Section
Molecular Biology Study Section (MBY)
Program Officer
Hackett, Charles J
Project Start
1999-01-01
Project End
2003-12-31
Budget Start
2001-01-01
Budget End
2001-12-31
Support Year
3
Fiscal Year
2001
Total Cost
$297,229
Indirect Cost
Name
Indiana University-Purdue University at Indianapolis
Department
Biochemistry
Type
Schools of Medicine
DUNS #
005436803
City
Indianapolis
State
IN
Country
United States
Zip Code
46202
Lockett, Angelia; Goebl, Mark G; Harrington, Maureen A (2008) Transient membrane recruitment of IRAK-1 in response to LPS and IL-1beta requires TNF R1. Am J Physiol Cell Physiol 295:C313-23
Kwon, Hyung-Joo; Breese, Erin Haag; Vig-Varga, Eva et al. (2004) Tumor necrosis factor alpha induction of NF-kappaB requires the novel coactivator SIMPL. Mol Cell Biol 24:9317-26
Yu, Kang-Yeol; Kwon, Hyung-Joo; Norman, David A M et al. (2002) Cutting edge: mouse pellino-2 modulates IL-1 and lipopolysaccharide signaling. J Immunol 169:4075-8
Vig, E; Green, M; Liu, Y et al. (2001) SIMPL is a tumor necrosis factor-specific regulator of nuclear factor-kappaB activity. J Biol Chem 276:7859-66
Green, M; Harrington, M A (2000) A comparison of macrophage colony-stimulating factor (M-CSF) gene expression in primary and immortalized endothelial cells. J Hematother Stem Cell Res 9:237-46
Vig, E; Green, M; Liu, Y et al. (1999) Modulation of tumor necrosis factor and interleukin-1-dependent NF-kappaB activity by mPLK/IRAK. J Biol Chem 274:13077-84
Pazdernik, N J; Donner, D B; Goebl, M G et al. (1999) Mouse receptor interacting protein 3 does not contain a caspase-recruiting or a death domain but induces apoptosis and activates NF-kappaB. Mol Cell Biol 19:6500-8