000095771 001__ 95771
000095771 005__ 20230921135433.0
000095771 0247_ $$2doi$$a10.3390/biom10091313
000095771 0248_ $$2sideral$$a120208
000095771 037__ $$aART-2020-120208
000095771 041__ $$aeng
000095771 100__ $$aNeira, J.L.
000095771 245__ $$aA phosphorylation-induced switch in the nuclear localization sequence of the intrinsically disordered nupr1 hampers binding to importin
000095771 260__ $$c2020
000095771 5060_ $$aAccess copy available to the general public$$fUnrestricted
000095771 5203_ $$aSeveral carrier proteins are involved in protein transport from the cytoplasm to the nucleus in eukaryotic cells. One of those is importin a, of which there are several human isoforms; among them, importin a3 (Impa3) has a high flexibility. The protein NUPR1, a nuclear protein involved in the cell-stress response and cell cycle regulation, is an intrinsically disordered protein (IDP) that has a nuclear localization sequence (NLS) to allow for nuclear translocation. NUPR1 does localize through the whole cell. In this work, we studied the affinity of the isolated wild-type NLS region (residues 54–74) of NUPR1 towards Impa3 and several mutants of the NLS region by using several biophysical techniques and molecular docking approaches. The NLS region of NUPR1 interacted with Impa3, opening the way to model the nuclear translocation of disordered proteins. All the isolated NLS peptides were disordered. They bound to Impa3 with low micromolar affinity (1.7–27 µM). Binding was hampered by removal of either Lys65 or Lys69 residues, indicating that positive charges were important; furthermore, binding decreased when Thr68 was phosphorylated. The peptide phosphorylated at Thr68, as well as four phospho-mimetic peptides (all containing the Thr68Glu mutation), showed the presence of a sequential NN(i, i + 1) nuclear Overhauser effect (NOE) in the 2D-1H-NMR (two-dimensional–proton NMR) spectra, indicating the presence of turn-like conformations. Thus, the phosphorylation of Thr68 modulates the binding of NUPR1 to Impa3 by a conformational, entropy-driven switch from a random-coil conformation to a turn-like structure.
000095771 536__ $$9info:eu-repo/grantAgreement/ES/DGA/B25-17R$$9info:eu-repo/grantAgreement/ES/DGA/E45-17R$$9info:eu-repo/grantAgreement/ES/ISCIII/CPII13-00017$$9info:eu-repo/grantAgreement/ES/ISCIII-ERDF-ESF/PI15-00663-Investing in your future$$9info:eu-repo/grantAgreement/ES/ISCIII-ERDF-ESF/PI18-00349-Investing in your future$$9info:eu-repo/grantAgreement/ES/MCIU-AEI-FEDER/BFU2016-78232-P$$9info:eu-repo/grantAgreement/ES/MCIU-AEI-FEDER/RTI2018-097991-B-I00
000095771 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000095771 590__ $$a4.879$$b2020
000095771 591__ $$aBIOCHEMISTRY & MOLECULAR BIOLOGY$$b96 / 296 = 0.324$$c2020$$dQ2$$eT1
000095771 592__ $$a1.125$$b2020
000095771 593__ $$aMolecular Biology$$c2020$$dQ2
000095771 593__ $$aBiochemistry$$c2020$$dQ2
000095771 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000095771 700__ $$aRizzuti, B.
000095771 700__ $$0(orcid)0000-0003-4726-7821$$aJiménez-Alesanco, A.$$uUniversidad de Zaragoza
000095771 700__ $$aPalomino-Schätzlein, M.
000095771 700__ $$0(orcid)0000-0001-5664-1729$$aAbián, O.$$uUniversidad de Zaragoza
000095771 700__ $$0(orcid)0000-0001-5702-4538$$aVelázquez-Campoy, A.$$uUniversidad de Zaragoza
000095771 700__ $$aIovanna, J.L.
000095771 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000095771 773__ $$g10, 9 (2020), 1313 [22 pp.]$$tBiomolecules$$x2218-273X
000095771 8564_ $$s637334$$uhttps://zaguan.unizar.es/record/95771/files/texto_completo.pdf$$yVersión publicada
000095771 8564_ $$s506640$$uhttps://zaguan.unizar.es/record/95771/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000095771 909CO $$ooai:zaguan.unizar.es:95771$$particulos$$pdriver
000095771 951__ $$a2023-09-21-13:30:34
000095771 980__ $$aARTICLE