Extended Data Fig. 1: Conservation and biochemical analysis of CeSPT-2.
From: The histone chaperone SPT2 regulates chromatin structure and function in Metazoa
a, Multiple protein sequence alignments corresponding to the three conserved regions found in metazoan SPT2 proteins are shown inside coloured boxes in red, yellow, and violet, respectively. Above the alignment that corresponds to the conserved SPT2 C-terminal region (in violet), the secondary structure of the experimentally determined H3-H4 HDB of human SPT2 (PDB: 5BS7 ref. 26) is shown (cylinders correspond to alpha helices). The amino acid colouring scheme indicates the average BLOSUM62 score (correlated to amino acid conservation) in each alignment column: black (greater than 3.5), grey (between 3.5 and 1.5) and light grey (between 1.5 and 0.5). Sequences are named according to their UniProt identifier, and species abbreviations are defined in the Supplementary Information file. b, Alignment of human and worm SPT2 histone binding domains. Red asterisks, residues in HsSPT2 reported to be required for the interaction with histones. Red arrow/box indicated the position of the Met residue histone binding defective mutation: M641 (human) / M627 (worm). c, d, Electrophoretic mobility shift assay (EMSA) showing binding of full-length recombinant CeSPT-2 or HsSPT2 (c), or of CeSPT-2 wild type and HBM (d), to synthetic cruciform DNA. Each arm of the four-way DNA junction contains 20 base pairs. For each panel, one representative experiment out of two is shown. e, Expression of GFP::CeSPT-2 WT or HBM in the indicated tissues of L4 larvae; one representative experiment of two. Images were taken with the same intensity and acquisition time.
