Fig. 2

Chromatin organization of the terminal TEL03L X element. a Schematic drawing of the TEL03L X element with the location of the HindIII site. The TEL03L-specific probe used in b and c is represented by a solid black line. TRF: Terminal Restriction Fragment. b In vivo ChEC experiments with GBD-MN (W3749-1a + pRSE), NLS-MN (W3749-1a + pG1NLS2), and MN-RAP1 (EPY007) analyzed on TEL03L. Southern blot with HindIII-digested genomic DNA hybridized to the TEL03L-specific probe. Time of MNase activity in minutes is indicated on top of gels. Solid line arrowheads show observable cutting common to all MNase-fused proteins, whereas the dotted line arrowhead corresponds to cutting observable predominately with MN-RAP1. M: DNA size marker with DNA marker sizes indicated on left of gel (kb). c Same as in b with H2A-MN (EPY130) and MN-RAP1 (EPY007) strains. The solid black arrowhead corresponds to very weak cutting observable with H2A-MN. d Location of MNase-sensitive sites on TEL03L with known features associated. Averaged fragment sizes ± standard deviation of MNase-fused protein-induced cutting determined from at least 2 independent experiments is plotted with respect to the HindIII site. Location of features (X-element, ACS, and Abf1, Tbf1- and Reb1-binding sites) is included. e Average distance ± standard deviation between MNase-sensitive sites identified in Fig. 2b, c. Size greater than 146 bp is congruent with positioning of a nucleosome, indicated by a dotted line. f Model of chromatin organization of TEL03L terminal fragment. Symbols as in Fig. 1, with addition of Abf1 as an oval