Sequence determinants of in cell condensate morphology, dynamics, and oligomerization as measured by number and brightness analysis

Cell Communication and Signaling

Lucia C. Strader¹  Alex S. Holehouse²  Ryan J. Emenecker³ 

[1]Center for Science and Engineering Living Systems (CSELS), Washington University, 63130, St. Louis, MO, USA

[2]Center for Engineering Mechanobiology, Washington University, 63130, St. Louis, MO, USA

[3]Department of Biology, Duke University, 27708, Durham, NC, USA

[4]Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, 63110, St. Louis, MO, USA

[5]Center for Science and Engineering Living Systems (CSELS), Washington University, 63130, St. Louis, MO, USA

[6]Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, 63110, St. Louis, MO, USA

[7]Center for Science and Engineering Living Systems (CSELS), Washington University, 63130, St. Louis, MO, USA

[8]Center for Engineering Mechanobiology, Washington University, 63130, St. Louis, MO, USA

关键词: Intrinsically disordered regions;    Biomolecular condensates;    Number and brightness analysis;    Fluorescence microscopy;    Fluorescence recovery after photobleaching;   

DOI  :  10.1186/s12964-021-00744-9

来源: Springer

【 摘 要 】

BackgroundBiomolecular condensates are non-stoichiometric assemblies that are characterized by their capacity to spatially concentrate biomolecules and play a key role in cellular organization. Proteins that drive the formation of biomolecular condensates frequently contain oligomerization domains and intrinsically disordered regions (IDRs), both of which can contribute multivalent interactions that drive higher-order assembly. Our understanding of the relative and temporal contribution of oligomerization domains and IDRs to the material properties of in vivo biomolecular condensates is limited. Similarly, the spatial and temporal dependence of protein oligomeric state inside condensates has been largely unexplored in vivo.MethodsIn this study, we combined quantitative microscopy with number and brightness analysis to investigate the aging, material properties, and protein oligomeric state of biomolecular condensates in vivo. Our work is focused on condensates formed by AUXIN RESPONSE FACTOR 19 (ARF19), a transcription factor integral to the auxin signaling pathway in plants. ARF19 contains a large central glutamine-rich IDR and a C-terminal Phox Bem1 (PB1) oligomerization domain and forms cytoplasmic condensates.ResultsOur results reveal that the IDR amino acid composition can influence the morphology and material properties of ARF19 condensates. In contrast the distribution of oligomeric species within condensates appears insensitive to the IDR composition. In addition, we identified a relationship between the abundance of higher- and lower-order oligomers within individual condensates and their apparent fluidity.ConclusionsIDR amino acid composition affects condensate morphology and material properties. In ARF condensates, altering the amino acid composition of the IDR did not greatly affect the oligomeric state of proteins within the condensate.87drinyczCdXeMm3m18s7MVideo Abstract

【 授权许可】

CC BY