Influence of histone H1 on the in vitro replication of DNA and chromatin

Abstract

The influence of histone H1 on DNA replication was studied using the SV40 in vitro replication system with two different templates: histone H1/DNA complexes and SV40 minichromosomes reconstituted with H1. We found that the cytosolic extracts used as a source of enzymes for in vitro replication contained high amounts of RNA which competed with template DNA for the binding of histone H1. Removal of this RNA was necessary to ensure the stability of the templates thus allowing for the first time the study of the replication of histone H1-carrying templates in vitro. In contrast to the inhibitory effect of histone H1 on the initiation of transcription, bound H1, when present at physiological ratios, does not interfere with the in vitro replication of DNA and minichromosomes. Ratios higher than one H1 molecule per nucleosome affected replication of reconstituted SV40 minichromosomes.