FRET Microscopy in Yeast

Overview

Journal Biosensors (Basel)

Specialty Biotechnology

Date 2019 Oct 17

PMID 31614546

Citations 12

Authors

Michal Skruzny

Emma Pohl

Marc Abella

Affiliations

Soon will be listed here.

Abstract

Förster resonance energy transfer (FRET) microscopy is a powerful fluorescence microscopy method to study the nanoscale organization of multiprotein assemblies in vivo. Moreover, many biochemical and biophysical processes can be followed by employing sophisticated FRET biosensors directly in living cells. Here, we summarize existing FRET experiments and biosensors applied in yeasts and , two important models of fundamental biomedical research and efficient platforms for analyses of bioactive molecules. We aim to provide a practical guide on suitable FRET techniques, fluorescent proteins, and experimental setups available for successful FRET experiments in yeasts.

Citing Articles

Transmembrane proteins in grape immunity: current knowledge and methodological advances.

Gallucci A, Giordano D, Facchiano A, Villano C, Carputo D, Aversano R Front Plant Sci. 2025; 15:1515163.

PMID: 39759230 PMC: 11695348. DOI: 10.3389/fpls.2024.1515163.

Cbf11 and Mga2 function together to activate transcription of lipid metabolism genes and promote mitotic fidelity in fission yeast.

Maresova A, Grulyova M, Hradilova M, Zemlianski V, Princova J, Prevorovsky M PLoS Genet. 2024; 20(12):e1011509.

PMID: 39652606 PMC: 11658701. DOI: 10.1371/journal.pgen.1011509.

Live cell microscopy: From image to insight.

Cuny A, Schlottmann F, Ewald J, Pelet S, Schmoller K Biophys Rev (Melville). 2024; 3(2):021302.

PMID: 38505412 PMC: 10903399. DOI: 10.1063/5.0082799.

Reticulons bind sphingolipids to activate the endoplasmic reticulum cell cycle checkpoint, the ER surveillance pathway.

Pina F, Yan B, Hu J, Niwa M Cell Rep. 2023; 42(12):113403.

PMID: 37979174 PMC: 11647836. DOI: 10.1016/j.celrep.2023.113403.

Genetically Encoded Fluorescent Probe for Detection of Heme-Induced Conformational Changes in Cytochrome c.

Genceroglu M, Cavdar C, Manioglu S, Bayraktar H Biosensors (Basel). 2023; 13(9).

PMID: 37754124 PMC: 10526477. DOI: 10.3390/bios13090890.

References

Tabke K, Albertmelcher A, Vitavska O, Huss M, Schmitz H, Wieczorek H . Reversible disassembly of the yeast V-ATPase revisited under in vivo conditions. Biochem J. 2014; 462(1):185-97. DOI: 10.1042/BJ20131293. View

Warren D, Andrews P, Gourlay C, Ayscough K . Sla1p couples the yeast endocytic machinery to proteins regulating actin dynamics. J Cell Sci. 2002; 115(Pt 8):1703-15. DOI: 10.1242/jcs.115.8.1703. View

Stoneman M, Paprocki J, Biener G, Yokoi K, Shevade A, Kuchin S . Quaternary structure of the yeast pheromone receptor Ste2 in living cells. Biochim Biophys Acta Biomembr. 2016; 1859(9 Pt A):1456-1464. DOI: 10.1016/j.bbamem.2016.12.008. View

Higuchi-Sanabria R, Garcia E, Tomoiaga D, Munteanu E, Feinstein P, Pon L . Characterization of Fluorescent Proteins for Three- and Four-Color Live-Cell Imaging in S. cerevisiae. PLoS One. 2016; 11(1):e0146120. PMC: 4699809. DOI: 10.1371/journal.pone.0146120. View

Teunissen A, Perez-Medina C, Meijerink A, Mulder W . Investigating supramolecular systems using Förster resonance energy transfer. Chem Soc Rev. 2018; 47(18):7027-7044. PMC: 6441672. DOI: 10.1039/c8cs00278a. View

Overton M, Blumer K . Use of fluorescence resonance energy transfer to analyze oligomerization of G-protein-coupled receptors expressed in yeast. Methods. 2002; 27(4):324-32. DOI: 10.1016/s1046-2023(02)00090-7. View

Young C, Raden D, Caplan J, Czymmek K, Robinson A . Cassette series designed for live-cell imaging of proteins and high-resolution techniques in yeast. Yeast. 2012; 29(3-4):119-36. PMC: 3371369. DOI: 10.1002/yea.2895. View

Schreiber G, Barberis M, Scolari S, Klaus C, Herrmann A, Klipp E . Unraveling interactions of cell cycle-regulating proteins Sic1 and B-type cyclins in living yeast cells: a FLIM-FRET approach. FASEB J. 2011; 26(2):546-54. DOI: 10.1096/fj.11-192518. View

Wegner S, Arslan H, Sunbul M, Yin J, He C . Dynamic copper(I) imaging in mammalian cells with a genetically encoded fluorescent copper(I) sensor. J Am Chem Soc. 2010; 132(8):2567-9. DOI: 10.1021/ja9097324. View

10.

Shin I, Ray J, Gupta V, Ilgu M, Beasley J, Bendickson L . Live-cell imaging of Pol II promoter activity to monitor gene expression with RNA IMAGEtag reporters. Nucleic Acids Res. 2014; 42(11):e90. PMC: 4066748. DOI: 10.1093/nar/gku297. View

11.

Jares-Erijman E, Jovin T . FRET imaging. Nat Biotechnol. 2003; 21(11):1387-95. DOI: 10.1038/nbt896. View

12.

Bermejo C, Haerizadeh F, Sadoine M, Chermak D, Frommer W . Differential regulation of glucose transport activity in yeast by specific cAMP signatures. Biochem J. 2013; 452(3):489-97. DOI: 10.1042/BJ20121736. View

13.

Bermejo C, Haerizadeh F, Takanaga H, Chermak D, Frommer W . Dynamic analysis of cytosolic glucose and ATP levels in yeast using optical sensors. Biochem J. 2010; 432(2):399-406. PMC: 2992555. DOI: 10.1042/BJ20100946. View

14.

Bajar B, Wang E, Zhang S, Lin M, Chu J . A Guide to Fluorescent Protein FRET Pairs. Sensors (Basel). 2016; 16(9). PMC: 5038762. DOI: 10.3390/s16091488. View

15.

Miller A, Chen J, Takasuka T, Jacobi J, Kaufman P, Irudayaraj J . Proliferating cell nuclear antigen (PCNA) is required for cell cycle-regulated silent chromatin on replicated and nonreplicated genes. J Biol Chem. 2010; 285(45):35142-54. PMC: 2966128. DOI: 10.1074/jbc.M110.166918. View

16.

Mastop M, Bindels D, Shaner N, Postma M, Gadella Jr T, Goedhart J . Characterization of a spectrally diverse set of fluorescent proteins as FRET acceptors for mTurquoise2. Sci Rep. 2017; 7(1):11999. PMC: 5607329. DOI: 10.1038/s41598-017-12212-x. View

17.

Jares-Erijman E, Jovin T . Imaging molecular interactions in living cells by FRET microscopy. Curr Opin Chem Biol. 2006; 10(5):409-16. DOI: 10.1016/j.cbpa.2006.08.021. View

18.

Bhaumik S . Analysis of in vivo targets of transcriptional activators by fluorescence resonance energy transfer. Methods. 2006; 40(4):353-9. DOI: 10.1016/j.ymeth.2006.06.025. View

19.

Skruzny M, Brach T, Ciuffa R, Rybina S, Wachsmuth M, Kaksonen M . Molecular basis for coupling the plasma membrane to the actin cytoskeleton during clathrin-mediated endocytosis. Proc Natl Acad Sci U S A. 2012; 109(38):E2533-42. PMC: 3458359. DOI: 10.1073/pnas.1207011109. View

20.

Peroza E, Ewald J, Parakkal G, Skotheim J, Zamboni N . A genetically encoded Förster resonance energy transfer sensor for monitoring in vivo trehalose-6-phosphate dynamics. Anal Biochem. 2015; 474:1-7. PMC: 4570562. DOI: 10.1016/j.ab.2014.12.019. View