Repression of histone gene transcription in quiescent 3T6 fibroblasts

Maintaining murine 3T6 fibroblasts in serum-depleted medium for a period of three days results in a resting cell population that does not synthesize DNA. Histone mRNA levels, closely tied to the cell-proliferation rate, are low due to a reduced rate of synthesis. A comparison of histone gene transcription in vitro by nuclear extracts of quiescent or proliferative 3T6 cells showed that a 200-bp segment of the promoter was responsible for repressing gene activity when cells were in a G0 state. In the absence of the distal promoter region (-200 to -400), gene transcription remained high in quiescent cells, indicating the proximal promoter region (+1 to -200) was responsible for basal gene activity. Alterations in protein binding to the distal promoter region correlated with histone H4 gene activity, suggesting that repression of histone gene transcription is linked to the attachment of a specific nuclear protein. During G1, the histone H4 gene was efficiently transcribed in vitro, but an inability to process the histone pre-mRNA limited the cellular content of mature histone mRNA. This distinction between transcriptional (in G0) and post-transcriptional (in G1) mechanisms for modulating histone mRNA levels suggests that gene-regulatory factors are specifically activated in quiescent cells to reduce expression of non-essential genes.