Structural Characteristics and Properties of the RNA-Binding Protein hnRNPK at Multiple Physical States

被引:0
作者
Le, Quang D. [1 ,2 ]
Lewis, Amanda [3 ]
Dix-Matthews, Alice [1 ]
Ringler, Philippe [3 ]
Duff, Anthony [4 ]
Whitten, Andrew E. [4 ]
Atkin, Rob [1 ]
Brunner, Manuel [1 ]
Ho, Diwei [1 ]
Iyer, K. Swaminathan [1 ]
Marshall, Andrew C. [1 ]
Fox, Archa H. [1 ,5 ]
Bond, Charles S. [1 ]
机构
[1] Univ Western Australia, Sch Mol Sci, Crawley, WA 6009, Australia
[2] VNU Univ Sci, Fac Biol, 334 Nguyen Trai St, Hanoi 100000, Vietnam
[3] Univ Basel, Ctr Cellular Imaging & NanoAnalyt C CINA, Biozentrum, CH-4001 Basel, Switzerland
[4] Australian Nucl Sci & Technol Org, Australian Ctr Neutron Scattering, New Illawarra Rd, Lucas Heights, NSW 2234, Australia
[5] Univ Western Australia, Sch Human Sci, Crawley, WA 6009, Australia
基金
英国医学研究理事会; 澳大利亚研究理事会; 澳大利亚国家健康与医学研究理事会;
关键词
hnRNPK protein; material state transitions; monomer; aggregate; phase separation; hydrogel; environmental factors; protein characteristics; LIQUID PHASE-SEPARATION; COMPLEXITY; AGGREGATION; DOMAINS; STRESS; METHYLATION;
D O I
10.3390/ijms26031356
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Heterogeneous nuclear ribonucleoprotein K (hnRNPK) is an RNA-binding protein containing low-complexity domains (LCDs), which are known to regulate protein behavior under stress conditions. This study demonstrates the ability to control hnRNPK's transitions into four distinct material states-monomer, soluble aggregate, liquid droplet, and fibrillar hydrogel-by modulating environmental factors such as temperature and protein concentration. Importantly, the phase-separated and hydrogel states are newly identified for eGFP-hnRNPK, marking a significant advancement in understanding its material properties. A combination of biophysical techniques, including DLS and SEC-LS, were used to further characterize hnRNPK in monomeric and soluble aggregate states. Structural methods, such as SANS, SAXS, and TEM, revealed the elongated morphology of the hnRNPK monomer. Environmental perturbations, such as decreased temperature or crowding agents, drove hnRNPK into phase-separated or gel-like states, each with distinct biophysical characteristics. These novel states were further analyzed using SEM, X-ray diffraction, and fluorescence microscopy. Collectively, these results demonstrate the complex behaviors of hnRNPK under different conditions and illustrate the properties of the protein in each material state. Transitions of hnRNPK upon condition changes could potentially affect functions of hnRNPK, playing a significant role in regulation of hnRNPK-involved processes in the cell.
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页数:22
相关论文
共 74 条
[1]   Considerations and Challenges in Studying Liquid-Liquid Phase Separation and Biomolecular Condensates [J].
Alberti, Simon ;
Gladfelter, Amy ;
Mittag, Tanja .
CELL, 2019, 176 (03) :419-434
[2]   A User's Guide for Phase Separation Assays with Purified Proteins [J].
Alberti, Simon ;
Saha, Shambaditya ;
Woodruff, Jeffrey B. ;
Franzmann, Titus M. ;
Wang, Jie ;
Hyman, Anthony A. .
JOURNAL OF MOLECULAR BIOLOGY, 2018, 430 (23) :4806-4820
[3]   The wisdom of crowds: regulating cell function through condensed states of living matter [J].
Alberti, Simon .
JOURNAL OF CELL SCIENCE, 2017, 130 (17) :2789-2796
[4]  
ARAKAWA T, 1985, METHOD ENZYMOL, V114, P49
[5]   X-ray crystallographic and NMR studies of the third KH domain of hnRNP K in complex with single-stranded nucleic acids [J].
Backe, PH ;
Messias, AC ;
Ravelli, RBG ;
Sattler, M ;
Cusack, S .
STRUCTURE, 2005, 13 (07) :1055-1067
[6]   In vivo aspects of protein folding and quality control [J].
Balchin, David ;
Hayer-Hartl, Manajit ;
Hartl, F. Ulrich .
SCIENCE, 2016, 353 (6294)
[7]   Biomolecular condensates: organizers of cellular biochemistry [J].
Banani, Salman F. ;
Lee, Hyun O. ;
Hyman, Anthony A. ;
Rosen, Michael K. .
NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2017, 18 (05) :285-298
[8]   Protein Phase Separation: A New Phase in Cell Biology [J].
Boeynaems, Steven ;
Alberti, Simon ;
Fawzi, Nicolas L. ;
Mittag, Tanja ;
Polymenidou, Magdalini ;
Rousseau, Frederic ;
Schymkowitz, Joost ;
Shorter, James ;
Wolozin, Benjamin ;
Van den Bosch, Ludo ;
Tompa, Peter ;
Fuxreiter, Monika .
TRENDS IN CELL BIOLOGY, 2018, 28 (06) :420-435
[9]   HnRNP K: One protein multiple processes [J].
Bomsztyk, K ;
Denisenko, O ;
Ostrowski, J .
BIOESSAYS, 2004, 26 (06) :629-638
[10]   LCD-Composer: an intuitive, composition-centric method enabling the identification and detailed functional mapping of low-complexity domains [J].
Cascarina, Sean M. ;
King, David C. ;
Nishimura, Erin Osborne ;
Ross, Eric D. .
NAR GENOMICS AND BIOINFORMATICS, 2021, 3 (02) :1-19