The major focus of the Fields laboratory has been the development and implementation of new technologies. Our motivation is that a novel technology can catalyze research across a spectrum of biological investigations, often leading to multiple applications beyond those initially envisioned. Technology development has led us to experimental questions that we had previously not explored, stimulated collaborations with computational biologists, biochemists and structural biologists, and provided opportunities to contribute to findings in medicine, as through studies of cancer-associated proteins, polyglutamine aggregation, aging, Toll-like receptors and malaria. Much of our research has centered on methods of protein analysis, although we have also put effort into methodologies to analyze DNA and RNA, to further synthetic biology approaches and to explore the engineering of bacteriophages.
For many of our technology efforts, we use the unicellular eukaryote Saccharomyces cerevisiae (baker's yeast) as the host organism for carrying out assays, but we also exploit E. coli, plant cells and tissue culture. Our philosophy on technology development is to pursue projects that can address important questions in basic biology or medicine and that can be readily applied by other labs. Recent projects have included the use of deep mutational scanning to analyze protein activities; the development of biosensors in bacteria and yeast; the identification of dominant negative mutants as reagents to inhibit protein function; and the use of mutant tRNAs to mistranslate proteins for genotype-phenotype studies.
Gamble, C.E., Brule, C.E., Dean, K.M., Fields, S. and Grayhack, E.J. (2016) Adjacent codons act in concert to modulate translation efficiency in yeast. Cell 166: 679-690.
Bhagavatula, G., Rich, M.S., Young, D., Marin, M. and Fields, S. (2017) A massively parallel fluorescence assay to detect the effects of synonymous mutations on TP53 expression. Molecular Cancer Research15: 1301-1307.
Cuperus, J.T, Groves, B., Kuchina, A., Rosenberg, A.B., Jojic, N., Fields, S. and Seelig, G. (2017) Deep learning of the regulatory grammar of yeast 5’ untranslated regions from 500,000 random sequences. Genome Research 27: 2015-2024.
Starita, L.M., Islam, M.M., Banerjee, T., Adamovich, A.I., Gullinsrud, J., Fields, S., Shendure, J. and Parvin, J.D. (2018) A multiplexed homology-directed DNA repair assay reveals the impact of more than 1,000 BRCA1 missense substitution variants on protein function. American Journal of Human Genetics 103: 498-508.
Brandsen, B.M., Mattheisen, J., Noel, T. and Fields, S. (2018) A biosensor strategy for E. coli based on ligand-dependent stabilization. ACS Synthetic Biology 7; 1990-1999.
Zimmerman, S.M., Kon, Y., Hauke, A.C., Ruiz, B.Y., Fields, S. and Phizicky, E.M. (2018) Conditional accumulation of toxic tRNAs to cause amino acid misincorporation. Nucleic Acids Research, 24: 410-422.
Dorrity, M.W., Queitsch, C. and Fields, S. (2019) High-throughput identification of dominant negative polypeptides in yeast. Nature Methods 16: 413-416.