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

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dc.contributor.advisorDiehl, Michael R
dc.creatorBellamkonda, Satya Kamala
dc.date.accessioned2021-04-13T22:06:20Z
dc.date.available2023-05-01T05:01:16Z
dc.date.created2021-05
dc.date.issued2021-03-26
dc.date.submittedMay 2021
dc.date.updated2021-04-13T22:06:21Z
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.
dc.embargo.terms2023-05-01
dc.format.mimetypeapplication/pdf
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>.
dc.identifier.urihttps://hdl.handle.net/1911/110268
dc.language.isoeng
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.
dc.subjectscaffold protein
dc.subjectsystems biology
dc.subjectsynthetic biology
dc.subjectimaging
dc.subjectflow cytometry
dc.subjectmammalian cell engineering
dc.subjectIQGAP1
dc.subjectprotein expression
dc.subjectsteady-state
dc.subjectprotein dynamics
dc.subjectPDL1
dc.titleInvestigations in Dynamic Protein Interactions and Steady-State Protein Abundance in Mammalian Cells
dc.typeThesis
dc.type.materialText
thesis.degree.departmentBioengineering
thesis.degree.disciplineEngineering
thesis.degree.grantorRice University
thesis.degree.levelDoctoral
thesis.degree.nameDoctor of Philosophy
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