Mapping the structure-activity landscape of ARHGAP36
Abstract/Contents
- Abstract
- ARHGAP36 is an atypical Rho GTPase-activating protein (GAP) family member that drives both spinal cord development and tumorigenesis, acting in part through an N-terminal motif that suppresses protein kinase A and activates Gli transcription factors. ARHGAP36 also contains isoform-specific N-terminal sequences, a central GAP-like module, and a unique C-terminal domain, and the functions of these regions remain unknown. In this dissertation, I describe our three-part approach to map the ARHGAP36 structure-activity landscape. Using splice variants and truncation mutant analyses, we identified an isoform-specific N-terminal sequence that inhibits ARHGAP36-mediated Gli activation and suppresses ARHGAP36 recruitment to the plasma membrane. The GAP-like and C-terminal domains can counter these inhibitory effects, in part by promoting trafficking to the plasma membrane and primary cilium, respectively. Through a high-throughput mutagenesis screen, we further discovered several residues in the GAP homology domain that are essential for Gli activation. Finally, we applied these variants in a comparative proteomic analysis that identified several GAP-like domain-dependent binding partners and demonstrated one interactor, prolyl oligopeptidase-like protein, to be a direct ARHGAP36 antagonist. In combination, these systems-level analyses have revealed an ensemble of mechanisms that could enable context-specific regulation of ARHGAP36 activity, establishing an experimental framework that can be applied towards other signaling proteins.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2021; ©2021 |
| Publication date | 2021; 2021 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Nano, Patricia Loren Rayos |
|---|---|
| Degree supervisor | Chen, James Kenneth |
| Thesis advisor | Chen, James Kenneth |
| Thesis advisor | Beachy, Philip Arden |
| Thesis advisor | Jackson, Peter |
| Thesis advisor | Meyer, Tobias |
| Degree committee member | Beachy, Philip Arden |
| Degree committee member | Jackson, Peter |
| Degree committee member | Meyer, Tobias |
| Associated with | Stanford University, Department of Chemical and Systems Biology |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Patricia Loren R. Nano. |
|---|---|
| Note | Submitted to the Department of Chemical and Systems Biology. |
| Thesis | Thesis Ph.D. Stanford University 2021. |
| Location | https://purl.stanford.edu/bp144bq0415 |
Access conditions
- Copyright
- © 2021 by Patricia Loren Rayos Nano
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