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. Author manuscript; available in PMC: 2010 Feb 10.
Published in final edited form as: Curr Biol. 2005 Nov 22;15(22):2044. doi: 10.1016/j.cub.2005.10.053

Figure 3.

Figure 3

The HIR complex and Asf1 assemble chromatin independently of DNA replication or CAF-1. (A) SV40 replication reactions containing α-32P-dATP. CAF-1 (10 ng, lane 2 or 20 ng, lane 3) or CBP-TAP purified HIR complex (2.25 μL, lanes 4 and 6 or 4.5 μL, lanes 5 and 7) were performed as indicated. Asf1/H3/H4 complexes (0.9 pmol) were added in lanes 6–8. Chromatin assembly was detected by the appearance of supercoiled DNA (indicated by arrow) on an agarose gel. Total DNA was visualized by staining with ethidium bromide (upper panel). Replicated molecules that incorporated α-32P-dATP were detected by autoradiography (lower panel). The values shown indicate the percentage of supercoiled DNA in each lane for both the total DNA and the replicated DNA. (B) The HIR complex does not stimulate replication dependent, CAF-1-mediated chromatin assembly. SV40 replication reactions were performed as in (A) in the presence of CAF-1 (7.5 ng, lanes 5–8, 10), histones H3/H4 (5 ng, lanes 2–4 and 6–8), HIR complex (4 μL, lanes 3, 7), HIR dialysis buffer (4 μL, lanes 4, 8), and pre-formed Asf1/H3/H4 complex (3.6 pmol, lanes 9, 10).