Abstract

In TR&D 1, we develop approaches to make complex changes to cells, and we generate biosensors to report on the effects of some of these changes. In the first aim, we establish strategies to rapidly and combinatorially up- or down-regulate a set of genes in a common pathway in order to improve the expression of a desired protein, and optimize these approaches using a vaccine protein that is commercially made in yeast. In the second aim, we devise new selections for use in chemostats and turbidostats to identify the effects of small changes in proteins, in promoters and in untranslated regions of mRNAs. Further, we modify the turbidostat platform to enable ramped selections, allowing feedback on temperature or drug concentration to optimize selections. In the third aim, we design new proteins that bind to a protein-protein interface and disrupt an interaction. These inhibitory proteins will allow us to assess the roles of individual members of large protein families that are difficult to analyze. Finally, in the fourth aim, we generate new biosensors to detect the levels of a small molecule and of a protein produced in yeast.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Biotechnology Resource Grants (P41)
Project #
2P41GM103533-21
Application #
9208071
Study Section
Special Emphasis Panel (ZRG1)
Project Start
1997-09-30
Project End
2022-03-31
Budget Start
2017-01-01
Budget End
2017-12-31
Support Year
21
Fiscal Year
2017
Total Cost
Indirect Cost

  Institution

Name
University of Washington
Department
Biochemistry
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195

  Publications

Tseng, Boo Shan; Reichhardt, Courtney; Merrihew, Gennifer E et al. (2018) A Biofilm Matrix-Associated Protease Inhibitor Protects Pseudomonas aeruginosa from Proteolytic Attack. MBio 9:
Yates 3rd, John R (2018) Content Is King: Databases Preserve the Collective Information of Science. J Biomol Tech 29:1-3
DaRosa, Paul A; Harrison, Joseph S; Zelter, Alex et al. (2018) A Bifunctional Role for the UHRF1 UBL Domain in the Control of Hemi-methylated DNA-Dependent Histone Ubiquitylation. Mol Cell 72:753-765.e6
Xu, Yi; Ju, Ho-Jong; DeBlasio, Stacy et al. (2018) A Stem-Loop Structure in Potato Leafroll Virus Open Reading Frame 5 (ORF5) Is Essential for Readthrough Translation of the Coat Protein ORF Stop Codon 700 Bases Upstream. J Virol 92:
Burke, Jordan E; Longhurst, Adam D; Merkurjev, Daria et al. (2018) Spliceosome Profiling Visualizes Operations of a Dynamic RNP at Nucleotide Resolution. Cell 173:1014-1030.e17
Xavier, Marina Amaral; Tirloni, Lucas; Pinto, Antônio F M et al. (2018) A proteomic insight into vitellogenesis during tick ovary maturation. Sci Rep 8:4698
Hollmann, Taylor; Kim, Tae Kwon; Tirloni, Lucas et al. (2018) Identification and characterization of proteins in the Amblyomma americanum tick cement cone. Int J Parasitol 48:211-224
Mutlu, Beste; Chen, Huei-Mei; Moresco, James J et al. (2018) Regulated nuclear accumulation of a histone methyltransferase times the onset of heterochromatin formation in C. elegans embryos. Sci Adv 4:eaat6224
Jester, Benjamin W; Tinberg, Christine E; Rich, Matthew S et al. (2018) Engineered Biosensors from Dimeric Ligand-Binding Domains. ACS Synth Biol 7:2457-2467
Kim, Yeoun Jin; Sweet, Steve M; Egertson, Jarrett D et al. (2018) Data-independent acquisition mass spectrometry to quantify protein levels in FFPE tumor biopsies for molecular diagnostics. J Proteome Res :

Showing the most recent 10 out of 372 publications