Open this publication in new window or tab >>Harvard Med Sch, Brigham & Womens Hosp, Div Rheumatol Immun & Inflammat, Boston, MA 02115 USA.;Harvard Med Sch, Harvard Med Sch Initiat RNA Med, Boston, MA 02115 USA..
Karolinska Inst, Dept Med Solna, Sci Life Lab, S-17177 Stockholm, Sweden.;Karolinska Univ Hosp, Div Infect Dis, S-17177 Stockholm, Sweden..
Karolinska Inst, Dept Microbiol Tumor & Cell Biol, S-17177 Stockholm, Sweden..
Karolinska Inst, Dept Microbiol Tumor & Cell Biol, S-17177 Stockholm, Sweden..
KTH, School of Biotechnology (BIO), Centres, Albanova VinnExcellence Center for Protein Technology, ProNova. KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Drug Discovery and Development.
Karolinska Inst, Dept Microbiol Tumor & Cell Biol, S-17177 Stockholm, Sweden..
Karolinska Inst, Dept Microbiol Tumor & Cell Biol, S-17177 Stockholm, Sweden..
KTH, School of Biotechnology (BIO), Centres, Albanova VinnExcellence Center for Protein Technology, ProNova. KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Protein Technology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
KTH, School of Biotechnology (BIO), Centres, Albanova VinnExcellence Center for Protein Technology, ProNova. KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Protein Engineering. KTH, Centres, Science for Life Laboratory, SciLifeLab.
KTH, School of Biotechnology (BIO), Centres, Albanova VinnExcellence Center for Protein Technology, ProNova. KTH, School of Engineering Sciences in Chemistry, Biotechnology and Health (CBH), Protein Science, Protein Engineering. KTH, Centres, Science for Life Laboratory, SciLifeLab.
Harvard Med Sch, Brigham & Womens Hosp, Div Rheumatol Immun & Inflammat, Boston, MA 02115 USA.;Harvard Med Sch, Harvard Med Sch Initiat RNA Med, Boston, MA 02115 USA..
Karolinska Inst, Dept Med Solna, Sci Life Lab, S-17177 Stockholm, Sweden.;Karolinska Univ Hosp, Div Infect Dis, S-17177 Stockholm, Sweden..
Karolinska Inst, Dept Microbiol Tumor & Cell Biol, S-17177 Stockholm, Sweden..
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2023 (English)In: Open Biology, E-ISSN 2046-2441, Vol. 13, no 5, article id 220369Article in journal (Refereed) Published
Abstract [en]
G3BP is the central node within stress-induced protein-RNA interaction networks known as stress granules (SGs). The SG-associated proteins Caprin-1 and USP10 bind mutually exclusively to the NTF2 domain of G3BP1, promoting and inhibiting SG formation, respectively. Herein, we present the crystal structure of G3BP1-NTF2 in complex with a Caprin-1-derived short linear motif (SLiM). Caprin-1 interacts with His-31 and His-62 within a third NTF2-binding site outside those covered by USP10, as confirmed using biochemical and biophysical-binding assays. Nano-differential scanning fluorimetry revealed reduced thermal stability of G3BP1-NTF2 at acidic pH. This destabilization was counterbalanced significantly better by bound USP10 than Caprin-1. The G3BP1/USP10 complex immunoprecipated from human U2OS cells was more resistant to acidic buffer washes than G3BP1/Caprin-1. Acidification of cellular condensates by approximately 0.5 units relative to the cytosol was detected by ratiometric fluorescence analysis of pHluorin2 fused to G3BP1. Cells expressing a Caprin-1/FGDF chimera with higher G3BP1-binding affinity had reduced Caprin-1 levels and slightly reduced condensate sizes. This unexpected finding may suggest that binding of the USP10-derived SLiM to NTF2 reduces the propensity of G3BP1 to enter condensates.
Place, publisher, year, edition, pages
The Royal Society, 2023
Keywords
stress granule, G3BP, caprin-1, crystal structure, condensate, pH
National Category
Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:kth:diva-328289 (URN)10.1098/rsob.220369 (DOI)000984747000003 ()37161291 (PubMedID)2-s2.0-85159669448 (Scopus ID)
Note
QC 20230607
2023-06-072023-06-072023-08-17Bibliographically approved