Fig. 2: Heterologous Dsup is nuclear localized in yeast and does not negatively impact growth. | Nature Communications

Fig. 2: Heterologous Dsup is nuclear localized in yeast and does not negatively impact growth.

From: Multivalent binding of the tardigrade Dsup protein to chromatin promotes yeast survival and longevity upon exposure to oxidative damage

Fig. 2

a Alignment of human HMGN1-3 identifies the HMGN core consensus (RRSARLSA). Also shown the R.varieornatus Dsup HMGN-like motif (aa 363-370, RRSSRLTS [underlined]) and alleles that mutate or delete this area and/or the downstream C-terminal region (aa 371-445) for phenotypic and/or biochemical studies (adapted from22) (Supplementary Data File 1E). Residues in red, including three functionally important arginines, are identical between the HMGN core consensus and Dsup. Green indicates the duplicated SV40 nuclear localization signal (NLS: PKKKRKVPKKKRKV) added to Dsup ∆HMGN ∆C for yeast expression. Pink indicates mutated residues in the HMGN-like motif to create -3R/3E (charge reversal) or -8A (charge neutralization). *, stop codon. b Schematic of Dsup wild-type (WT) and mutant alleles stably expressed in yeast for phenotypic studies (Figs. 1–4 [Dsup ∆C + NLS (as used in Fig. 7) not depicted]). N-terminal 6xHIS and C-terminal FLAG tags on each protein are not depicted. c Immunofluorescence to examine subcellular location of Dsup alleles (anti-FLAG) in yeast. DNA stain DAPI identifies nuclei. H2A-FLAG and GAPDH are respective controls for nuclear and cytoplasmic localization. EV, Empty vector. d Immunoblot shows relative expression of Dsup alleles (anti-FLAG) in yeast. Anti-GAPDH is a loading control from samples run on the same gel. e Representative growth curves of yeast expressing Dsup alleles. Source data are provided as a Source Data file.

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