The FERM-PTP Pez is a multifunctional protein that has previously been shown to play a role in epithelial/limb development, actin organization, and tissue growth, and has recently been identified as an upstream regulator of the Hippo pathway restricting stem cell proliferation. This study used molecular and genetic techniques to elucidate Pez interactions and function at the membrane. In order to determine the membrane binding partner of Pez, we tested various phospholipids in a protein:lipid overlay experiment. We established phosphatidylserine as the primary candidate for lipid-FERM interaction. Additionally, we found that Pez has the ability to dimerize through sites within the Central Linker. In constructs lacking the FERM domain, the Central Linker-PTP portion of Pez undergoes organizational changes that result in membrane sheets that coalesce into stable cytoplasmic filaments that can be isolated from tissues. To determine if Pez filaments can incorporate membrane or other proteins during its transformation from a membrane sheet, we co-expressed Pez filaments with membrane markers or known interactors and determined filaments contain membrane markers and can encapsulate potential binding partners.
To identify Pez binding partners, we conducted a affinity purification/mass spectrometry experiment using the FERM-CL portion of Pez, and found two main groups of potential interactors; Hippo pathway components and cytoskeletal proteins. We focused on the Hippo pathway interactors and found Pez 1) does not phenocopy effects of starvation, 2) genetically interacts with the tumor suppressor Warts, 3) has a progressive role in maintaining midgut homeostasis, and 4) is a direct negative regulator of Yki.
|Advisor:||Edwards, Kevin A.|
|Commitee:||Friesen, Jona A., Galewsky, Samuel, Kirik, Viktor M., Larson, Erik D.|
|School:||Illinois State University|
|Department:||Molecular and Cellular Biology|
|School Location:||United States -- Illinois|
|Source:||DAI-B 74/12(E), Dissertation Abstracts International|
|Subjects:||Molecular biology, Cellular biology, Oncology|
|Keywords:||Hippo, Pez, Protein tyrosine phosphatase, Tumor suppressor, Yorkie|
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