This proposal examines a novel protein kinase called PKK which associates with protein kinase C beta which can activate NF kappa B, and which is required for the activation of NF kappa B by phorbol ester. PKK is also expressed at very high levels in some human breast cancer cell lines.
One aim i s to understand how PKK is activated in terms of ligands, receptors, and upstream activators. PKK is heavily phosphorylated and the role of PKK phosphorylation in catalytic activation and the ability of PKK to activate NFkappa B will be explored in detail. Since certain MAP3 Kinases interact with PKK, a possible role of these MAP3Ks as upstream activators of PKK will be explored. The identity of putative upstream activating kinases that may participate in the phosphorylation of a critical activation loop residue of PKK will also be sought. The identification of upstream activators will narrow the search for possible ligands and pathways involved in PKK activation in a number of cell types. Specific dominant negative forms of PKK and the use of stable siRNA constructs will facilitate these studies.
A second aim i s to understand how PKK activates NF kappa B. The structural features of PKK that contribute to NFkappa B activation will be examined in detail using site-directed mutagenesis. Our studies will explore how PKK interacts directly or indirectly with components of the IKK complex. Finally we will explore whether PKK contributes to NFkappa B activation in breast cancer and whether PKK is required for tumorigenesis. The potential role of PKK in mammary epithelial differentiation will be explored and PKK's role in mammary development will be analyzed using a conditional knockout approach. PKK represents a novel enzyme that may be critical in pathways that are important in cancer, inflammation, cell survival, and differentiation.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA102793-04
Application #
7098095
Study Section
Chemical Pathology Study Section (CPA)
Program Officer
Yassin, Rihab R,
Project Start
2003-09-24
Project End
2008-05-31
Budget Start
2006-06-01
Budget End
2007-05-31
Support Year
4
Fiscal Year
2006
Total Cost
$363,191
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
073130411
City
Boston
State
MA
Country
United States
Zip Code
02199
Cariappa, Annaiah; Boboila, Cristian; Moran, Stewart T et al. (2007) The recirculating B cell pool contains two functionally distinct, long-lived, posttransitional, follicular B cell populations. J Immunol 179:2270-81
Cariappa, Annaiah; Chase, Catharine; Liu, Haoyuan et al. (2007) Naive recirculating B cells mature simultaneously in the spleen and bone marrow. Blood 109:2339-45
Moran, Stewart T; Cariappa, Annaiah; Liu, Haoyuan et al. (2006) Protein kinase C-associated kinase is not required for the development of peripheral B lymphocyte populations. Mol Immunol 43:1694-9
Cariappa, Annaiah; Mazo, Irina B; Chase, Catharine et al. (2005) Perisinusoidal B cells in the bone marrow participate in T-independent responses to blood-borne microbes. Immunity 23:397-407
Cariappa, Annaiah; Shoham, Tsipi; Liu, Haoyuan et al. (2005) The CD9 tetraspanin is not required for the development of peripheral B cells or for humoral immunity. J Immunol 175:2925-30
Pillai, Shiv; Cariappa, Annaiah; Moran, Stewart T (2005) Marginal zone B cells. Annu Rev Immunol 23:161-96
Pillai, Shiv; Cariappa, Annaiah; Moran, Stewart T (2004) Positive selection and lineage commitment during peripheral B-lymphocyte development. Immunol Rev 197:206-18