Biomolecular polyelectrolyte complexes can be formed between oppositely charged intrinsically disordered regions (IDRs) of proteins or between IDRs and nucleic acids. Highly charged IDRs are abundant in the nucleus, yet few have been functionally characterized. Here, we show that a positively charged IDR within the human ATP-dependent DNA helicase Q4 (RECQ4) forms coacervates with G-quadruplexes (G4s). We describe a three-step model of charge-driven coacervation by integrating equilibrium and kinetic binding data in a global numerical model. The oppositely charged IDR and G4 molecules form a complex in the solution that follows a rapid nucleation-growth mechanism leading to a dynamic equilibrium between dilute and condensed phases. We also discover a physical interaction with Replication Protein A (RPA) and demonstrate that the IDR can switch between the two extremes of the structural continuum of complexes. The structural, kinetic, and thermodynamic profile of its interactions revealed a dynamic disordered complex with nucleic acids and a static ordered complex with RPA protein. The two mutually exclusive binding modes suggest a regulatory role for the IDR in RECQ4 function by enabling molecular handoffs. Our study extends the functional repertoire of IDRs and demonstrates a role of polyelectrolyte complexes involved in G4 binding.

CEITEC Central European Institute of Technology Masaryk University Brno Czech Republic

Department of Biology Faculty of Medicine Masaryk University Brno Czech Republic

Department of Condensed Matter Physics Faculty of Science Masaryk University Brno Czech Republic

Institute of Structural and Molecular Biology University College London London WC1E 6BT UK

International Clinical Research Center St Anne's University Hospital Brno Czech Republic

Loschmidt Laboratories Department of Experimental Biology and RECETOX Faculty of Science Masaryk University Brno Czech Republic

National Centre for Biomolecular Research Faculty of Science Masaryk University Brno Czech Republic

School of Pharmacy University College London London WC1N 1AX UK

See more in PubMed

Bochman ML, Paeschke K, Zakian VA. DNA secondary structures: stability and function of G-quadruplex structures. Nat. Rev. Genet. 2012;13:770–780. PubMed PMC

Burge S, Parkinson GN, Hazel P, Todd AK, Neidle S. Quadruplex DNA: sequence, topology and structure. Nucleic Acids Res. 2006;34:5402–5415. PubMed PMC

Williamson JR, Raghuraman MK, Cech TR. Monovalent cation-induced structure of telomeric DNA: the G-quartet model. Cell. 1989;59:871–880. PubMed

Sfen D, Gilbert W. Formation of parallel four-stranded complexes by guanine-rich motif for meiosis. Nature. 1998;334:364–366. PubMed

Spiegel J, Adhikari S, Balasubramanian S. The structure and function of DNA G-quadruplexes. Trends Chem. 2020;2:123–136. PubMed PMC

Prorok P, et al. Involvement of G-quadruplex regions in mammalian replication origin activity. Nat. Commun. 2019;10:3274. PubMed PMC

Valton AL, et al. G4 motifs affect origin positioning and efficiency in two vertebrate replicators. EMBO J. 2014;33:732–746. PubMed PMC

Huang WC, et al. Direct evidence of mitochondrial G-quadruplex DNA by using fluorescent anti-cancer agents. Nucleic Acids Res. 2015;43:10102–10113. PubMed PMC

Koirala D, et al. Intramolecular folding in three tandem guanine repeats of human telomeric DNA. Chem. Commun. 2012;48:2006–2008,. PubMed

Siddiqui-Jain A, Grand CL, Bearss DJ, Hurley LH. Direct evidence for a G-quadruplex in a promoter region and its targeting with a small molecule to repress c-MYC transcription. Proc. Natl Acad. Sci. USA. 2002;99:11593–11598. PubMed PMC

Balasubramanian S, Hurley LH, Neidle S. Targeting G-quadruplexes in gene promoters: a novel anticancer strategy? Nat. Rev. Drug Discov. 2011;10:261–275. PubMed PMC

Yuan L, et al. Existence of G-quadruplex structures in promoter region of oncogenes confirmed by G-quadruplex DNA cross-linking strategy. Sci. Rep. 2013;3:1811. PubMed PMC

Varshney D, Spiegel J, Zyner K, Tannahill D, Balasubramanian S. The regulation and functions of DNA and RNA G-quadruplexes. Nat. Rev. Mol. Cell Biol. 2020;21:459–474. PubMed PMC

Besnard E, et al. Unraveling cell type-specific and reprogrammable human replication origin signatures associated with G-quadruplex consensus motifs. Nat. Struct. Mol. Biol. 2012;19:837–844. PubMed

Lerner LK, Sale JE. Replication of G Quadruplex DNA. Genes. 2019;10:95–95. PubMed PMC

Maffia, A., Ranise, C. & Sabbioneda, S. From R-loops to G-quadruplexes: emerging new threats for the replication fork. Int. J. Mol. Sci.21, 1506 (2020). PubMed PMC

Lee WTC, et al. Single-molecule imaging reveals replication fork coupled formation of G-quadruplex structures hinders local replication stress signaling. Nat. Commun. 2021;12:2525. PubMed PMC

Sarkies P, Reams C, Simpson LJ, Sale JE. Epigenetic instability due to defective replication of structured DNA. Mol. Cell. 2010;40:703–713. PubMed PMC

Mao SQ, et al. DNA G-quadruplex structures mold the DNA methylome. Nat. Struct. Mol. Biol. 2018;25:951–957. PubMed PMC

Reina C, Cavalieri V. Epigenetic modulation of chromatin states and gene expression by G-quadruplex structures. Int. J. Mol. Sci. 2020;21:1–22. PubMed PMC

Paeschke K, Simonsson T, Postberg J, Rhodes D, Lipps HJ. Telomere end-binding proteins control the formation of G-quadruplex DNA structures in vivo. Nat. Struct. Mol. Biol. 2005;12:847–854. PubMed

Hansel-Hertsch R, et al. G-quadruplex structures mark human regulatory chromatin. Nat. Genet. 2016;48:1267–1272. PubMed

Sun H, Karow JK, Hickson ID, Maizels N. The Bloom’s syndrome helicase unwinds G4 DNA. J. Biol. Chem. 1998;273:27587–27592. PubMed

Sun H, Bennett RJ, Maizels N. The Saccharomyces cerevisiae Sgs1 helicase efficiently unwinds G-G paired DNAs. Nucleic Acids Res. 1999;27:1978–1984. PubMed PMC

Fry M, Loeb LA. Human werner syndrome DNA helicase unwinds tetrahelical structures of the fragile X syndrome repeat sequence d(CGG)n. J. Biol. Chem. 1999;274:12797–12802. PubMed

Wu X, Maizels N. Substrate-specific inhibition of RecQ helicase. Nucleic Acids Res. 2001;29:1765–1771. PubMed PMC

Wu Y, Shin-ya K, Brosh RM., Jr FANCJ helicase defective in Fanconia anemia and breast cancer unwinds G-quadruplex DNA to defend genomic stability. Mol. Cell Biol. 2008;28:4116–4128. PubMed PMC

Bharti SK, et al. Specialization among iron-sulfur cluster helicases to resolve G-quadruplex DNA structures that threaten genomic stability. J. Biol. Chem. 2013;288:28217–28229. PubMed PMC

Wu CG, Spies M. G-quadruplex recognition and remodeling by the FANCJ helicase. Nucleic Acids Res. 2016;44:8742–8753. PubMed PMC

Vaughn JP, et al. The DEXH protein product of the DHX36 gene is the major source of tetramolecular quadruplex G4-DNA resolving activity in HeLa cell lysates. J. Biol. Chem. 2005;280:38117–38120. PubMed

Sauer M, et al. DHX36 prevents the accumulation of translationally inactive mRNAs with G4-structures in untranslated regions. Nat. Commun. 2019;10:2421. PubMed PMC

Paeschke K, Capra JA, Zakian VA. DNA replication through G-quadruplex motifs is promoted by the Saccharomyces cerevisiae Pif1 DNA helicase. Cell. 2011;145:678–691. PubMed PMC

Paeschke K, et al. Pif1 family helicases suppress genome instability at G-quadruplex motifs. Nature. 2013;497:458–462. PubMed PMC

Wu L, Hickson ID. DNA helicases required for homologous recombination and repair of damaged replication forks. Annu. Rev. Genet. 2006;40:279–306. PubMed

Tippana R, Hwang H, Opresko PL, Bohr VA, Myong S. Single-molecule imaging reveals a common mechanism shared by G-quadruplex-resolving helicases. Proc. Natl Acad. Sci. USA. 2016;113:8448–8453. PubMed PMC

Chen WF, et al. Molecular mechanistic insights into Drosophila DHX36-mediated G-quadruplex unfolding: a structure-based model. Structure. 2018;26:403–415.e404. PubMed

Chen MC, et al. Structural basis of G-quadruplex unfolding by the DEAH/RHA helicase DHX36. Nature. 2018;558:465–469. PubMed PMC

Voter AF, Qiu Y, Tippana R, Myong S, Keck JL. A guanine-flipping and sequestration mechanism for G-quadruplex unwinding by RecQ helicases. Nat. Commun. 2018;9:4201. PubMed PMC

Qureshi MH, Ray S, Sewell AL, Basu S, Balci H. Replication protein A unfolds G-quadruplex structures with varying degrees of efficiency. J. Phys. Chem. B. 2012;116:5588–5594. PubMed PMC

Safa L, et al. Binding polarity of RPA to telomeric sequences and influence of G-quadruplex stability. Biochimie. 2014;103:80–88. PubMed

Safa L, et al. 5’ to 3’ Unfolding directionality of DNA secondary structures by replication protein A: G-quadruplexes and duplexes. J. Biol. Chem. 2016;291:21246–21256. PubMed PMC

Wu W, et al. HERC2 facilitates BLM and WRN helicase complex interaction with RPA to suppress G-quadruplex DNA. Cancer Res. 2018;78:6371–6385. PubMed

Maestroni L, et al. RPA and Pif1 cooperate to remove G-rich structures at both leading and lagging strand. Cell Stress. 2020;4:48–63. PubMed PMC

Wang B, et al. Liquid-liquid phase separation in human health and diseases. Signal Transduct. Target Ther. 2021;6:290. PubMed PMC

Weis K, Hondele M. The role of DEAD-box ATPases in gene expression and the regulation of RNA-protein condensates. Annu. Rev. Biochem. 2022;91:197–219. PubMed

Wang T, et al. Bloom syndrome helicase compresses single-stranded DNA into phase-separated condensates. Angew. Chem. Int. Ed. Engl. 2022;61:e202209463. PubMed

Kaiser S, Sauer F, Kisker C. The structural and functional characterization of human RecQ4 reveals insights into its helicase mechanism. Nat. Commun. 2017;8:15907. PubMed PMC

Masumoto H, Muramatsu S, Kamimura Y, Araki H. S-Cdk-dependent phosphorylation of Sld2 essential for chromosomal DNA replication in budding yeast. Nature. 2002;415:651–655. PubMed

Tanaka S, et al. CDK-dependent phosphorylation of Sld2 and Sld3 initiates DNA replication in budding yeast. Nature. 2007;445:328–332. PubMed

Zegerman P, Diffley JF. Phosphorylation of Sld2 and Sld3 by cyclin-dependent kinases promotes DNA replication in budding yeast. Nature. 2007;445:281–285. PubMed

Gaggioli V, et al. CDK phosphorylation of SLD-2 is required for replication initiation and germline development in C. elegans. J. Cell. Biol. 2014;204:507–522. PubMed PMC

Sangrithi MN, et al. Initiation of DNA replication requires the RECQL4 protein mutated in Rothmund-Thomson syndrome. Cell. 2005;121:887–898. PubMed

Matsuno K, Kumano M, Kubota Y, Hashimoto Y, Takisawa H. The N-terminal noncatalytic region of Xenopus RecQ4 is required for chromatin binding of DNA polymerase alpha in the initiation of DNA replication. Mol. Cell Biol. 2006;26:4843–4852. PubMed PMC

Abe T, et al. The N-terminal region of RECQL4 lacking the helicase domain is both essential and sufficient for the viability of vertebrate cells. Role of the N-terminal region of RECQL4 in cells. Biochim. Biophys. Acta. 2011;1813:473–479. PubMed

Castillo-Tandazo W, et al. ATP-dependent helicase activity is dispensable for the physiological functions of Recql4. PLoS Genet. 2019;15:e1008266. PubMed PMC

Sedlackova H, Cechova B, Mlcouskova J, Krejci L. RECQ4 selectively recognizes Holliday junctions. DNA Repair. 2015;30:80–89. PubMed

Keller H, et al. The intrinsically disordered amino-terminal region of human RecQL4: multiple DNA-binding domains confer annealing, strand exchange and G4 DNA binding. Nucleic Acids Res. 2014;42:12614–12627. PubMed PMC

Im JS, et al. Assembly of the Cdc45-Mcm2-7-GINS complex in human cells requires the Ctf4/And-1, RecQL4, and Mcm10 proteins. Proc. Natl Acad. Sci. USA. 2009;106:15628–15632. PubMed PMC

Xu X, Rochette PJ, Feyissa EA, Su TV, Liu Y. MCM10 mediates RECQ4 association with MCM2-7 helicase complex during DNA replication. EMBO J. 2009;28:3005–3014. PubMed PMC

Kliszczak M, et al. Interaction of RECQ4 and MCM10 is important for efficient DNA replication origin firing in human cells. Oncotarget. 2015;6:40464–40479. PubMed PMC

Lu H, et al. Cell cycle-dependent phosphorylation regulates RECQL4 pathway choice and ubiquitination in DNA double-strand break repair. Nat. Commun. 2017;8:2039. PubMed PMC

Lu H, et al. RECQL4 promotes DNA end resection in repair of DNA double-strand breaks. Cell Rep. 2016;16:161–173. PubMed PMC

Shamanna RA, et al. RECQ helicase RECQL4 participates in non-homologous end joining and interacts with the Ku complex. Carcinogenesis. 2014;35:2415–2424. PubMed PMC

De S, et al. RECQL4 is essential for the transport of p53 to mitochondria in normal human cells in the absence of exogenous stress. J. Cell Sci. 2012;125:2509–2522. PubMed

Ohlenschlager O, et al. The N-terminus of the human RecQL4 helicase is a homeodomain-like DNA interaction motif. Nucleic Acids Res. 2012;40:8309–8324. PubMed PMC

Nguyen Ba AN, et al. Proteome-wide discovery of evolutionary conserved sequences in disordered regions. Sci. Signal. 2012;5:rs1. PubMed PMC

Davey NE, et al. Attributes of short linear motifs. Mol. Biosyst. 2012;8:268–281. PubMed

Zarin T, et al. Proteome-wide signatures of function in highly diverged intrinsically disordered regions. Elife. 2019;8:1–26. PubMed PMC

Uversky VN. Natively unfolded proteins: a point where biology waits for physics. Protein Sci. 2002;11:739–756. PubMed PMC

Mittag T, Kay LE, Forman-Kay JD. Protein dynamics and conformational disorder in molecular recognition. J. Mol. Recognit. 2010;23:105–116. PubMed

Berlow RB, Dyson HJ, Wright PE. Functional advantages of dynamic protein disorder. FEBS Lett. 2015;589:2433–2440. PubMed PMC

Csizmok V, Follis AV, Kriwacki RW, Forman-Kay JD. Dynamic protein interaction networks and new structural paradigms in signaling. Chem. Rev. 2016;116:6424–6462. PubMed PMC

Dunker AK, Cortese MS, Romero P, Iakoucheva LM, Uversky VN. Flexible nets. The roles of intrinsic disorder in protein interaction networks. FEBS J. 2005;272:5129–5148. PubMed

Fuxreiter M, et al. Malleable machines take shape in eukaryotic transcriptional regulation. Nat. Chem. Biol. 2008;4:728–737. PubMed PMC

Oldfield CJ, Dunker AK. Intrinsically disordered proteins and intrinsically disordered protein regions. Annu. Rev. Biochem. 2014;83:553–584. PubMed

Borgia A, et al. Extreme disorder in an ultrahigh-affinity protein complex. Nature. 2018;555:61–66. PubMed PMC

Aramburu IV, Lemke EA. Floppy but not sloppy: Interaction mechanism of FG-nucleoporins and nuclear transport receptors. Semin. Cell Dev. Biol. 2017;68:34–41. PubMed PMC

Fung HYJ, Birol M, Rhoades E. IDPs in macromolecular complexes: the roles of multivalent interactions in diverse assemblies. Curr. Opin. Struct. Biol. 2018;49:36–43. PubMed PMC

Tan PS, et al. Two differential binding mechanisms of FG-nucleoporins and nuclear transport receptors. Cell Rep. 2018;22:3660–3671. PubMed PMC

Sottini A, et al. Polyelectrolyte interactions enable rapid association and dissociation in high-affinity disordered protein complexes. Nat. Commun. 2020;11:5736. PubMed PMC

Uversky VN. Protein intrinsic disorder-based liquid-liquid phase transitions in biological systems: complex coacervates and membrane-less organelles. Adv. Colloid Interface Sci. 2017;239:97–114. PubMed

Uversky VN. Intrinsically disordered proteins in overcrowded milieu: membrane-less organelles, phase separation, and intrinsic disorder. Curr. Opin. Struct. Biol. 2017;44:18–30. PubMed

Turner AL, et al. Highly disordered histone H1-DNA model complexes and their condensates. Proc. Natl Acad. Sci. USA. 2018;115:11964–11969. PubMed PMC

Banani SF, Lee HO, Hyman AA, Rosen MK. Biomolecular condensates: organizers of cellular biochemistry. Nat. Rev. Mol. Cell Biol. 2017;18:285–298. PubMed PMC

Das RK, Ruff KM, Pappu RV. Relating sequence encoded information to form and function of intrinsically disordered proteins. Curr. Opin. Struct. Biol. 2015;32:102–112. PubMed PMC

Vuzman D, Levy Y. DNA search efficiency is modulated by charge composition and distribution in the intrinsically disordered tail. Proc. Natl Acad. Sci. USA. 2010;107:21004–21009. PubMed PMC

Brosh RM, Jr, et al. Functional and physical interaction between WRN helicase and human replication protein A. J. Biol. Chem. 1999;274:18341–18350. PubMed

Brosh RM, Jr, et al. Replication protein A physically interacts with the Bloom’s syndrome protein and stimulates its helicase activity. J. Biol. Chem. 2000;275:23500–23508. PubMed

Doherty KM, et al. Physical and functional mapping of the replication protein a interaction domain of the werner and bloom syndrome helicases. J. Biol. Chem. 2005;280:29494–29505. PubMed

Kang D, et al. Interaction of replication protein A with two acidic peptides from human Bloom syndrome protein. FEBS Lett. 2018;592:547–558. PubMed

Bythell-Douglas R, Deans AJ. A structural guide to the Bloom syndrome complex. Structure. 2021;29:99–113. PubMed

Marechal A, Zou L. RPA-coated single-stranded DNA as a platform for post-translational modifications in the DNA damage response. Cell Res. 2015;25:9–23. PubMed PMC

Sugitani N, Chazin WJ. Characteristics and concepts of dynamic hub proteins in DNA processing machinery from studies of RPA. Prog. Biophys. Mol. Biol. 2015;117:206–211. PubMed PMC

Mer G, et al. Structural basis for the recognition of DNA repair proteins UNG2, XPA, and RAD52 by replication factor RPA. Cell. 2000;103:449–456. PubMed

Ali SI, Shin JS, Bae SH, Kim B, Choi BS. Replication protein A 32 interacts through a similar binding interface with TIPIN, XPA, and UNG2. Int. J. Biochem. Cell Biol. 2010;42:1210–1215. PubMed

Xie S, et al. Structure of RPA32 bound to the N-terminus of SMARCAL1 redefines the binding interface between RPA32 and its interacting proteins. FEBS J. 2014;281:3382–3396. PubMed

Zhao W, et al. Promotion of BRCA2-dependent homologous recombination by DSS1 via RPA targeting and DNA mimicry. Mol. Cell. 2015;59:176–187. PubMed PMC

Marino F, et al. Structural and biochemical characterization of an RNA/DNA binding motif in the N-terminal domain of RecQ4 helicases. Sci. Rep. 2016;6:21501. PubMed PMC

Luu KN, Phan AT, Kuryavyi V, Lacroix L, Patel DJ. Structure of the human telomere in K+ solution: an intramolecular (3 + 1) G-quadruplex scaffold. J. Am. Chem. Soc. 2006;128:9963–9970. PubMed PMC

Adrian M, et al. Structure and conformational dynamics of a stacked dimeric G-quadruplex formed by the human CEB1 minisatellite. J. Am. Chem. Soc. 2014;136:6297–6305. PubMed

Kriwacki RW, Hengst L, Tennant L, Reed SI, Wright PE. Structural studies of p21Waf1/Cip1/Sdi1 in the free and Cdk2-bound state: conformational disorder mediates binding diversity. Proc. Natl Acad. Sci. USA. 1996;93:11504–11509. PubMed PMC

Tompa P, Fuxreiter M. Fuzzy complexes: polymorphism and structural disorder in protein-protein interactions. Trends Biochem. Sci. 2008;33:2–8. PubMed

Berjanskii MV, Wishart DS. The RCI server: rapid and accurate calculation of protein flexibility using chemical shifts. Nucleic Acids Res. 2007;35:W531–W537. PubMed PMC

Hafsa NE, Wishart DS. CSI 2.0: a significantly improved version of the Chemical Shift Index. J. Biomol. NMR. 2014;60:131–146. PubMed

Shen Y, Bax A. Protein backbone and sidechain torsion angles predicted from NMR chemical shifts using artificial neural networks. J. Biomol. NMR. 2013;56:227–241. PubMed PMC

Peng JW, Wagner G. Mapping of spectral density-functions using heteronuclear NMR relaxation measurements. J Magn Reson. 1992;98:308–332.

Peng JW, Wagner G. Mapping of the spectral densities of N-H bond motions in eglin c using heteronuclear relaxation experiments. Biochemistry. 1992;31:8571–8586. PubMed

Waudby CA, Ramos A, Cabrita LD, Christodoulou J. Two-dimensional NMR lineshape analysis. Sci. Rep. 2016;6:24826. PubMed PMC

Waudby CA, Christodoulou J. NMR lineshape analysis of intrinsically disordered protein interactions. Methods Mol. Biol. 2020;2141:477–504. PubMed PMC

Kim JY, Meng F, Yoo J, Chung HS. Diffusion-limited association of disordered protein by non-native electrostatic interactions. Nat. Commun. 2018;9:4707. PubMed PMC

Tzeng SR, Kalodimos CG. Dynamic activation of an allosteric regulatory protein. Nature. 2009;462:368–372. PubMed

Amaral M, et al. Protein conformational flexibility modulates kinetics and thermodynamics of drug binding. Nat. Commun. 2017;8:2276. PubMed PMC

Mendoza O, Bourdoncle A, Boule JB, Brosh RM, Jr., Mergny JL. G-quadruplexes and helicases. Nucleic Acids Res. 2016;44:1989–2006. PubMed PMC

Do NQ, Phan AT. Monomer-dimer equilibrium for the 5’-5’ stacking of propeller-type parallel-stranded G-quadruplexes: NMR structural study. Chemistry. 2012;18:14752–14759. PubMed

Bolognesi B, et al. A concentration-dependent liquid phase separation can cause toxicity upon increased protein expression. Cell Rep. 2016;16:222–231. PubMed PMC

Sing CE, Perry SL. Recent progress in the science of complex coacervation. Soft Matter. 2020;16:2885–2914. PubMed

Del Villar-Guerra R, Trent JO, Chaires JB. G-quadruplex secondary structure obtained from circular dichroism spectroscopy. Angew. Chem. Int. Ed. Engl. 2018;57:7171–7175. PubMed PMC

Yevdokimov YM, et al. Re-entrant cholesteric phase in DNA liquid-crystalline dispersion particles. J. Biol. Phys. 2017;43:45–68. PubMed PMC

Mimura M, et al. Quadruplex folding promotes the condensation of linker histones and DNAs via liquid-liquid phase separation. J. Am. Chem. Soc. 2021;143:9849–9857. PubMed

Renaud de la Faverie A, Guedin A, Bedrat A, Yatsunyk LA, Mergny JL. Thioflavin T as a fluorescence light-up probe for G4 formation. Nucleic Acids Res. 2014;42:e65. PubMed PMC

Xu S, et al. Thioflavin T as an efficient fluorescence sensor for selective recognition of RNA G-quadruplexes. Sci. Rep. 2016;6:24793. PubMed PMC

Shen Y, et al. Biomolecular condensates undergo a generic shear-mediated liquid-to-solid transition. Nat. Nanotechnol. 2020;15:841–847. PubMed PMC

London TB, et al. FANCJ is a structure-specific DNA helicase associated with the maintenance of genomic G/C tracts. J. Biol. Chem. 2008;283:36132–36139. PubMed PMC

Chang LW, et al. Sequence and entropy-based control of complex coacervates. Nat. Commun. 2017;8:1273. PubMed PMC

Zervoudis NA, Obermeyer AC. The effects of protein charge patterning on complex coacervation. Soft Matter. 2021;17:6637–6645. PubMed

Vieregg JR, et al. Oligonucleotide-peptide complexes: phase control by hybridization. J. Am. Chem. Soc. 2018;140:1632–1638. PubMed

Flock T, Weatheritt RJ, Latysheva NS, Babu MM. Controlling entropy to tune the functions of intrinsically disordered regions. Curr. Opin. Struct. Biol. 2014;26:62–72. PubMed

Wright PE, Dyson HJ. Intrinsically disordered proteins in cellular signalling and regulation. Nat. Rev. Mol. Cell Biol. 2015;16:18–29. PubMed PMC

Shin G, Jeong D, Kim H, Im JS, Lee JK. RecQL4 tethering on the pre-replicative complex induces unscheduled origin activation and replication stress in human cells. J. Biol. Chem. 2019;294:16255–16265. PubMed PMC

Takai H, Smogorzewska A, de Lange T. DNA damage foci at dysfunctional telomeres. Curr. Biol. 2003;13:1549–1556. PubMed

Ghosh AK, et al. RECQL4, the protein mutated in Rothmund-Thomson syndrome, functions in telomere maintenance. J. Biol. Chem. 2012;287:196–209. PubMed PMC

Croteau DL, et al. RECQL4 localizes to mitochondria and preserves mitochondrial DNA integrity. Aging Cell. 2012;11:456–466. PubMed PMC

Kumari J, et al. Mitochondrial functions of RECQL4 are required for the prevention of aerobic glycolysis-dependent cell invasion. J. Cell Sci. 2016;129:1312–1318. PubMed

Petkovic M, Dietschy T, Freire R, Jiao R, Stagljar I. The human Rothmund-Thomson syndrome gene product, RECQL4, localizes to distinct nuclear foci that coincide with proteins involved in the maintenance of genome stability. J. Cell Sci. 2005;118:4261–4269. PubMed

Singh DK, et al. The involvement of human RECQL4 in DNA double-strand break repair. Aging Cell. 2010;9:358–371. PubMed PMC

Tsuyama T, et al. N-terminal region of RecQ4 inhibits non-homologous end joining and chromatin association of the Ku heterodimer in Xenopus egg extracts. Gene. 2021;787:145647. PubMed

Falabella M, Fernandez RJ, Johnson FB, Kaufman BA. Potential roles for G-quadruplexes in mitochondria. Curr. Med. Chem. 2019;26:2918–2932. PubMed PMC

Alberti S, Gladfelter A, Mittag T. Considerations and challenges in studying liquid-liquid phase separation and biomolecular condensates. Cell. 2019;176:419–434. PubMed PMC

Ghodgaonkar MM, et al. Phenotypic characterization of missense polymerase-delta mutations using an inducible protein-replacement system. Nat. Commun. 2014;5:4990. PubMed

Henricksen LA, Umbricht CB, Wold MS. Recombinant replication protein A: expression, complex formation, and functional characterization. J. Biol. Chem. 1994;269:11121–11132. PubMed

Sattler M, Schleucher J, Griesinger C. Heteronuclear multidimensional NMR experiments for the structure determination of proteins in solution employing pulsed field gradients. Prog. Nucl. Mag. Res. Sp. 1999;34:93–158.

Evangelidis T, et al. Automated NMR resonance assignments and structure determination using a minimal set of 4D spectra. Nat. Commun. 2018;9:384. PubMed PMC

Lee W, Tonelli M, Markley JL. NMRFAM-SPARKY: enhanced software for biomolecular NMR spectroscopy. Bioinformatics. 2015;31:1325–1327. PubMed PMC

Korzhnev DM, Billeter M, Arseniev AS, Orekhov VY. NMR studies of Brownian tumbling and internal motions in proteins. Prog. Nucl. Mag. Res. Sp. 2001;38:197–266.

Eaton, J. W., Bateman, D., Hauberg, S. & Wehbring, R. GNU Octave version 3.8.1 manual: a high-level interactive language for numerical computations. CreateSpace Independent Publishing Platform. (2014). ISBN 1441413006, http://www.gnu.org/software/octave/doc/interpreter/.

Ishima R, Nagayama K. Protein backbone dynamics revealed by quasi spectral density function analysis of amide N-15 nuclei. Biochemistry. 1995;34:3162–3171. PubMed

Ishima R, Nagayama K. Quasi-spectral-density function analysis for nitrogen-15 nuclei in proteins. J. Magn. Reson. 1995;108:73–76.

Farrow NA, Zhang O, Szabo A, Torchia DA, Kay LE. Spectral density function mapping using 15N relaxation data exclusively. J. Biomol. NMR. 1995;6:153–162. PubMed

Ferrage F, Cowburn D, Ghose R. Accurate sampling of high-frequency motions in proteins by steady-state (15)N-{(1)H} nuclear Overhauser effect measurements in the presence of cross-correlated relaxation. J. Am. Chem. Soc. 2009;131:6048–6049. PubMed PMC

Johnson KA, Simpson ZB, Blom T. Global kinetic explorer: a new computer program for dynamic simulation and fitting of kinetic data. Anal. Biochem. 2009;387:20–29. PubMed

Li A, Ziehr JL, Johnson KA. A new general method for simultaneous fitting of temperature and concentration dependence of reaction rates yields kinetic and thermodynamic parameters for HIV reverse transcriptase specificity. J. Biol. Chem. 2017;292:6695–6702. PubMed PMC

RECQ4-MUS81 interaction contributes to telomere maintenance with implications to Rothmund-Thomson syndrome