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Fig. 2. Increases in histone mRNA levels in vivo and YY1-histone alpha-element-binding activity in vitro upon entry into S phase. (A) Total RNA was isolated from stable transfectant CHO cells for each time interval indicated across the top (AS, asynchronous; numbers are hours after mitotic shake-off). A fragment of the mouse H3.2-614 gene (3'-end-labelled) was used as a radioactive probe in S1-nuclease protection assays, with 3 µg of total RNA per reaction. The positions of the probe fragments protected by the mouse and hamster histone transcripts (arrows) and the positions of markers (nucleotide number) are indicated. (B) An electrophoretic mobility-shift assay using the duplex histone alpha oligonucleotides as a probe was carried out with nuclear extracts from synchronous populations of CHO cells (lanes 0, 1, 3, 5 and 7.5; numbers are hours after shake-off) or from mouse myeloma cells (lane MM) as a positive control. The position of the complex (resulting from YY1 DNA-binding activity) and free probe are indicated.

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