Colon cancer is a major health challenge, ranking as the second leading cause of cancer deaths. Most cases are initiated by mutations in the tumor suppressor Adenomatous polyposis coli (APC). APC is best known as a key negative regulator of the Wnt cell-cell signaling pathway, which shapes cell fates in virtually every tissue and organ, regulates homeostasis of stem cells in several tissues, and is inappropriately activated in many cancers. APC also plays Wnt-independent roles in regulating the cytoskeleton, thus facilitating processes like high-fidelity chromosome segregation, which also are disrupted in cancer. Our lab's long term goal is to determine how APC and its protein partners regulate both Wnt signaling and the cytoskeleton during normal development and homeostasis, and how that process goes wrong in colon and other cancers. Our lab played important roles in defining how APC acts in the "destruction complex", which targets the Wnt effectors sscatenin for ubiquitination and destruction, and also identified APC as a regulator of cytoskeletal events. To do so, we established powerful dual model systems to study this in parallel in intact animals during Drosophila development and in cultured human cells, using a highly multidisciplinary approach combining state-of-the-art genetic, cell biological, and biochemical tools. Understanding mechanisms by which APC and its protein partners regulate both Wnt signaling and the cytoskeleton during normal development will thus advance both basic science and clinical applications. While we now possess textbook models for both Wnt regulation and APCs cytoskeletal function, it is becoming increasingly clear that these paradigms are oversimplified-in fact APC's mechanistic roles in the destruction complex and in cytoskeletal regulation remain largely mysterious. Our work in the past funding period challenged existing hypotheses for APC function in Wnt regulation, including our own, demonstrating that APC does not play essential nuclear roles in Wnt regulation nor does it play an essential role in localizing the destruction complex. Instead, work from our lab and others suggests the innovative hypothesis underlying Aim 1, in which APC regulates a dynamic catalytic cycle of the destruction complex, which is essential for sscatenin transfer to the E3 ubiquitin ligase.
In Aim 1 we will define mechanisms by which APC and Axin act in a dynamic destruction complex, identify novel protein partners that regulate the cycle of conformational change, and determine how Wnt signaling alters destruction complex dynamics to turn it off. Our data also suggest textbook views of APC's roles as a cytoskeletal regulator must be significantly modified. We hypothesize APC is not an essential regulator of mitosis, but instead acts to ensure high fidelity chromosome segregation, working as part of a multiprotein complex, and with its action buffered by mitotic checkpoints that can partially compensate for its loss-this underlies Aim 2. In this Aim we will determine how APC and its partners act to regulate centrosome migration and thus ensure high fidelity chromosome segregation, and explore how Chk2 and other checkpoint regulators buffer the errors that occur in its absence.
Most cases of colon cancer begin with mutations in the APC gene. We will define APC's roles in communication between cells, and in maintaining the stability of the genome through its effects on the cell's skeleton, thus providing the insights needed to develop more effective treatments.
|Poulton, John S; Cuningham, John C; Peifer, Mark (2014) Acentrosomal Drosophila epithelial cells exhibit abnormal cell division, leading to cell death and compensatory proliferation. Dev Cell 30:731-45|
|Poulton, John S; Mu, Frank W; Roberts, David M et al. (2013) APC2 and Axin promote mitotic fidelity by facilitating centrosome separation and cytoskeletal regulation. Development 140:4226-36|
|Roberts, David M; Pronobis, Mira I; Alexandre, Kelly M et al. (2012) Defining components of the ýý-catenin destruction complex and exploring its regulation and mechanisms of action during development. PLoS One 7:e31284|
|Roberts, David M; Pronobis, Mira I; Poulton, John S et al. (2012) Regulation of Wnt signaling by the tumor suppressor adenomatous polyposis coli does not require the ability to enter the nucleus or a particular cytoplasmic localization. Mol Biol Cell 23:2041-56|
|Roberts, David M; Pronobis, Mira I; Poulton, John S et al. (2011) Deconstructing the ßcatenin destruction complex: mechanistic roles for the tumor suppressor APC in regulating Wnt signaling. Mol Biol Cell 22:1845-63|
|Cadigan, Ken M; Peifer, Mark (2009) Wnt signaling from development to disease: insights from model systems. Cold Spring Harb Perspect Biol 1:a002881|
|Rogers, Gregory C; Rusan, Nasser M; Roberts, David M et al. (2009) The SCF Slimb ubiquitin ligase regulates Plk4/Sak levels to block centriole reduplication. J Cell Biol 184:225-39|
|Rusan, Nasser M; Peifer, Mark (2008) Original CIN: reviewing roles for APC in chromosome instability. J Cell Biol 181:719-26|
|Rogers, Gregory C; Rusan, Nasser M; Peifer, Mark et al. (2008) A multicomponent assembly pathway contributes to the formation of acentrosomal microtubule arrays in interphase Drosophila cells. Mol Biol Cell 19:3163-78|
|Rusan, Nasser M; Akong, Kathryn; Peifer, Mark (2008) Putting the model to the test: are APC proteins essential for neuronal polarity, axon outgrowth, and axon targeting? J Cell Biol 183:203-12|
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