Protomer alignment modulates specificity of RNA substrate recognition by Ire1.

Autor:	Li W; Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, United States.; Howard Hughes Medical Institute, San Francisco, United States., Crotty K; Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, United States.; Howard Hughes Medical Institute, San Francisco, United States., Garrido Ruiz D; Department of Pharmaceutical Chemistry, University of California at San Francisco, San Francisco, United States., Voorhies M; Department of Microbiology and Immunology, University of California at San Francisco, San Francisco, United States., Rivera C; Department of Molecular Biophysics and Biochemistry, Yale School of Medicine, New Haven, United States., Sil A; Department of Microbiology and Immunology, University of California at San Francisco, San Francisco, United States., Mullins RD; Howard Hughes Medical Institute, San Francisco, United States.; Department of Cellular and Molecular Pharmacology, University of California at San Francisco, San Francisco, United States., Jacobson MP; Department of Pharmaceutical Chemistry, University of California at San Francisco, San Francisco, United States., Peschek J; Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, United States.; Howard Hughes Medical Institute, San Francisco, United States., Walter P; Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, United States.; Howard Hughes Medical Institute, San Francisco, United States.
Jazyk:	angličtina
Zdroj:	ELife [Elife] 2021 Apr 27; Vol. 10. Date of Electronic Publication: 2021 Apr 27.
DOI:	10.7554/eLife.67425
Abstrakt:	The unfolded protein response (UPR) maintains protein folding homeostasis in the endoplasmic reticulum (ER). In metazoan cells, the Ire1 branch of the UPR initiates two functional outputs-non-conventional mRNA splicing and selective mRNA decay (RIDD). By contrast, Ire1 orthologs from Saccharomyces cerevisiae and Schizosaccharomyces pombe are specialized for only splicing or RIDD, respectively. Previously, we showed that the functional specialization lies in Ire1's RNase activity, which is either stringently splice-site specific or promiscuous (Li et al., 2018). Here, we developed an assay that reports on Ire1's RNase promiscuity. We found that conversion of two amino acids within the RNase domain of S. cerevisiae Ire1 to their S. pombe counterparts rendered it promiscuous. Using biochemical assays and computational modeling, we show that the mutations rewired a pair of salt bridges at Ire1 RNase domain's dimer interface, changing its protomer alignment. Thus, Ire1 protomer alignment affects its substrates specificity. Competing Interests: WL, KC, DG, MV, CR, AS, RM, MJ, JP, PW No competing interests declared (© 2021, Li et al.)
Databáze:	MEDLINE
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