The tardigrade damage suppressor protein binds to nucleosomes and protects DNA from hydroxyl radicals
- University of California, San Diego, United States
Figures
Figure 1
R.varieornatus damage suppressor protein (Rv Dsup).
(A) Rv Dsup is a highly charged protein. The protein was analyzed by using the MPI Bioinformatics Toolkit (Zimmermann et al., 2018), and it is predicted to be disordered except for some potentially …
Figure 2 with 1 supplement
Rv Dsup binds preferentially to mononucleosomes relative to free DNA.
(A) Gel mobility shift analysis with the Xenopus borealis 5S rDNA (147 bp) as either mononucleosomes or free DNA. The indicated amounts of purified Rv Dsup was combined with the mononucleosomes or …
Figure 2—figure supplement 1
Quantitation of gel mobility shift analyses of Rv Dsup binding to mononucleosomes or free DNA.
Gel mobility shift experiments were performed with Rv Dsup as in Figure 2 of the main text. For each sample, the percent of the mononucleosomes or free DNA that was bound by Dsup was determined by …
Figure 3
Rv Dsup can be incorporated into periodic nucleosome arrays.
The ATP-dependent assembly of periodic nucleosome arrays was carried out with purified ACF, dNLP, core histones, ATP, relaxed plasmid DNA, and topoisomerase I (Fyodorov and Kadonaga, 2003; Khuong et …
Figure 4
Rv Dsup and histone H1 can bind simultaneously to nucleosomes.
(A) Native gel mobility shift analysis reveals the binding of both Rv Dsup and histone H1 to mononucleosomes. Experiments were performed with 5S rDNA mononucleosomes (181 bp DNA) and the indicated …
Figure 5
The Dsup-like protein from the tardigrade Hypsibius exemplaris is a nucleosome-binding protein.
(A) Comparison of the genomic regions in the vicinity of the genes encoding R. varieornatus Dsup (Rv Dsup) and H. exemplaris Dsup-like protein (He Dsup). This diagram is based on sequences from R. …
Figure 6
Dsup protects chromatin from hydroxyl radical-mediated DNA cleavage.
Hydroxyl radical-mediated cleavage of DNA was carried out with plasmid pGIE-0 (3.3 kb) as either free DNA or chromatin in the absence or presence of the indicated factors. Where noted, hydroxyl …
Figure 7
A region of sequence similarity between Dsup proteins and the nucleosome-binding domain of vertebrate HMGN proteins is important for the binding of Dsup to nucleosomes.
(A) Alignment of Dsup proteins with the five human HMGN proteins as representative members of the HMGN protein family. Partial sequences of Rv Dsup, He Dsup, and the five human HMGN proteins are …
Additional files
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- https://doi.org/10.7554/eLife.47682.011
