Polycystic kidney disease (PKD) is one of the most common lethal genetic diseases. Mutations in the Sterile Alpha Motif (SAM) domains of Bicaudal-C (Bicc1) and Samcystin/Anks6 are known to cause cystic disease in humans, rats or mice by unknown mechanisms. Prior work, and our own preliminary results, have established an interaction network between the SAM domains of Bicc1, Anks6 and a new protein we have identified called Anks3. Bicc1 regulates cell polarity and the expression of several proteins known to contribute to PKD. The fact that both Anks3 and Anks6 bind to the key regulatory protein Bicc1, suggests that they may be important for Bicc1 function. We propose to test two primary hypotheses: (1) That Anks6 and Anks3 can modulate the functions of Bicc1 either separately or together via there SAM domain interactions. (2) That the newly identified Anks3 protein can modulate the development of PKD.
Aim 1. We will physically and structurally characterize the SAM mediated interactions of Bicc1, Ank3 and Anks6. An understanding of the architecture of the complexes will be important for understanding the biological consequences of SAM domain mutations.
Aim 2. Test the hypothesis that Anks3 and Anks6 modulate known Bicc1 functions in cellular assays. We will examine the effects of Anks3 an Anks6 on cellular localization of Bicc1, protein expression and Wnt signaling.
Aim 3. Test the hypothesis that Anks3 mutations can generate PKD in transgenic rats. Transgenic rat strains will be created where Anks3 is over-expressed or deleted and examined for the development of cystic disease. We will also study how these alterations affect the development of PKD in Anks6 mutant rats. The project has the potential to explain the mechanism of several disease-causing mutations, identify functions of proteins involved in PKD, and identify a new player in the complex pathway to PKD.

Public Health Relevance

Polycystic kidney disease (PKD) is one of the most common lethal genetic diseases. The disease is characterized by the development of fluid filled cysts in the kidney and ultimately renal failure. This project seeks to understand how mutations in two interacting proteins, called Bicaudal-C and SamCystin cause cystic disease. The investigators have also discovered a third protein, called Anks3, that binds to both Bicaudal-C and SamCystin. They therefore plan to investigate the involvement of Anks3 in the genesis of cystic disease. The project has the potential to describe a new pathway to PKD, establish a new animal model for PKD, and identify a new therapeutic target.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK100482-04
Application #
9205230
Study Section
Kidney Molecular Biology and Genitourinary Organ Development (KMBD)
Program Officer
Maric-Bilkan, Christine
Project Start
2014-01-01
Project End
2018-12-31
Budget Start
2017-01-01
Budget End
2018-12-31
Support Year
4
Fiscal Year
2017
Total Cost
Indirect Cost
Name
University of California Los Angeles
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Rothé, Benjamin; Leettola, Catherine N; Leal-Esteban, Lucia et al. (2018) Crystal Structure of Bicc1 SAM Polymer and Mapping of Interactions between the Ciliopathy-Associated Proteins Bicc1, ANKS3, and ANKS6. Structure 26:209-224.e6
Jefferson, Robert E; Min, Duyoung; Corin, Karolina et al. (2018) Applications of Single-Molecule Methods to Membrane Protein Folding Studies. J Mol Biol 430:424-437
Min, Duyoung; Arbing, Mark A; Jefferson, Robert E et al. (2016) A simple DNA handle attachment method for single molecule mechanical manipulation experiments. Protein Sci 25:1535-44
Delestré, Laure; Bakey, Zeineb; Prado, Cécilia et al. (2015) ANKS3 Co-Localises with ANKS6 in Mouse Renal Cilia and Is Associated with Vasopressin Signaling and Apoptosis In Vivo in Mice. PLoS One 10:e0136781
Leettola, Catherine N; Knight, Mary Jane; Cascio, Duilio et al. (2014) Characterization of the SAM domain of the PKD-related protein ANKS6 and its interaction with ANKS3. BMC Struct Biol 14:17