Investigations in Dynamic Protein Interactions and Steady-State Protein Abundance in Mammalian Cells

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dc.contributor.advisorDiehl, Michael Ren_US
dc.creatorBellamkonda, Satya Kamalaen_US
dc.date.accessioned2021-04-13T22:06:20Zen_US
dc.date.available2023-05-01T05:01:16Zen_US
dc.date.created2021-05en_US
dc.date.issued2021-03-26en_US
dc.date.submittedMay 2021en_US
dc.date.updated2021-04-13T22:06:21Zen_US
dc.description.abstractThe coordination of lipid messenger signaling with cytoskeletal regulation is central to many organelle-specific regulatory processes. Multi-domain scaffold proteins orchestrate the activities of multiple signaling intermediates and regulatory proteins on organelles. Investigating scaffold functions is challenging because these interactions occur at different timescales that are not well understood through standard static colocalization analyses. This work employs live-cell imaging to probe how the multi-domain scaffold IQGAP1 coordinates the activities of proteins affecting local actin polymerization, membrane processing, and phosphoinositide signaling, and point to a scaffold tethering mechanism. This work also focuses on engineering mammalian cells to modulate steady-state protein expression. Protein abundance is a highly regulated cellular process, and variations in protein levels leads to changes in cellular phenotypes and functions. These changes may cause human genetic diseases and changes in immune response, and it is important to build tunable mammalian cell libraries that can be used for systematic analyses to study the link between protein expression levels and cellular function. In this project, mammalian cell libraries with subpopulations of variable steady-state expression of a fluorescent protein or membrane ligand were made by multiplexing synthetic biology strategies in a modular, plug-and-play, high-throughput system. These libraries generated cell lines with tunable, stable, robust expression with quick turnaround times, which can be employed for future functional analyses of protein expression levels on cellular activity.en_US
dc.embargo.terms2023-05-01en_US
dc.format.mimetypeapplication/pdfen_US
dc.identifier.citationBellamkonda, Satya Kamala. "Investigations in Dynamic Protein Interactions and Steady-State Protein Abundance in Mammalian Cells." (2021) Diss., Rice University. <a href="https://hdl.handle.net/1911/110268">https://hdl.handle.net/1911/110268</a>.en_US
dc.identifier.urihttps://hdl.handle.net/1911/110268en_US
dc.language.isoengen_US
dc.rightsCopyright is held by the author, unless otherwise indicated. Permission to reuse, publish, or reproduce the work beyond the bounds of fair use or other exemptions to copyright law must be obtained from the copyright holder.en_US
dc.subjectscaffold proteinen_US
dc.subjectsystems biologyen_US
dc.subjectsynthetic biologyen_US
dc.subjectimagingen_US
dc.subjectflow cytometryen_US
dc.subjectmammalian cell engineeringen_US
dc.subjectIQGAP1en_US
dc.subjectprotein expressionen_US
dc.subjectsteady-stateen_US
dc.subjectprotein dynamicsen_US
dc.subjectPDL1en_US
dc.titleInvestigations in Dynamic Protein Interactions and Steady-State Protein Abundance in Mammalian Cellsen_US
dc.typeThesisen_US
dc.type.materialTexten_US
thesis.degree.departmentBioengineeringen_US
thesis.degree.disciplineEngineeringen_US
thesis.degree.grantorRice Universityen_US
thesis.degree.levelDoctoralen_US
thesis.degree.nameDoctor of Philosophyen_US
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