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. 2020 Sep 30;219(11):e202007135. doi: 10.1083/jcb.202007135

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© 2020 MRC Laboratory of Molecular Biology

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure 6. — Sec31 disordered region can be functionally replaced with unrelated sequences. (A) Serial dilutions of the indicated constructs where the yeast Sec31 disordered region was replaced with that of other species reveals that only the human Sec31A disordered region can support viability. (B) In vitro microsomal budding experiments using the indicated proteins correlates with viability; only the Hs31A fusion can support vesicle formation with both GTP and GMP-PNP. (C) Serial dilutions of the indicated strains reveals that deletion of the active fragment, or mutation of the PPP or WN motifs within the Hs31A disordered region reduces viability. (D) Serial dilutions of Sec31-Sec16 disordered region chimera reveals that only the first Sec16 disordered region can support viability when replacing the Sec31_DR. (E) In vitro budding from microsomal membranes with the indicated proteins correlates with viability, with only Nhis-Sec31-16_DR1 budding. (F) Serial dilutions of chimeric Sec31 constructs containing the indicated unrelated disordered regions reveals that the Las17 domain swap supports viability.