bioRxiv preprint doi: https://doi.org/10.1101/771592; this version posted September 16, 2019. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 Arginine-enriched mixed-charge domains provide cohesion for nuclear 2 speckle condensation 3 4 Jamie A. Greig1, Tu Anh Nguyen1, Michelle Lee1, Alex S. Holehouse2, Ammon E. 5 Posey2, Rohit V. Pappu2, and Gregory JeDD1* 6 7 1Temasek Life Sciences Laboratory & Department of Biological Sciences, The 8 National University of Singapore, Singapore 117604 9 2Department of BiomeDical Engineering anD Center for Science & Engineering of 10 Living Systems (CSELS), Washington University in St. Louis, St. Louis, MO 63130, 11 USA 12 13 *CorresponDence:
[email protected] 14 15 Abstract 16 Low-complexity protein Domains promote the formation of various biomolecular 17 conDensates. However, in many cases, the precise sequence features governing 18 conDensate formation anD iDentity remain unclear. Here, we investigate the role of 19 intrinsically DisordereD mixeD-charge Domains (MCDs) in nuclear speckle 20 conDensation. Proteins composeD exclusively of arginine/aspartic-aciD DipeptiDe 21 repeats unDergo length-DepenDent conDensation anD speckle incorporation. 22 Substituting arginine with lysine in synthetic anD natural speckle-associateD MCDs 23 abolishes these activities, iDentifying a key role for multivalent contacts through 24 arginine’s guaniDinium ion. MCDs can synergise with a speckle-associateD RNA 25 recognition motif to promote speckle specificity anD resiDence. MCD behaviour is 26 tuneable through net-charge: increasing negative charge abolishes conDensation anD 27 speckle incorporation.