Histone-Gene Reiteration in the Genome of Mouse

Abstract

Histone messenger RNA labeled to high specific activity was isolated and purified from the mouse tissue-culture cell line L929. It had an electrophoretic mobility equivalent to a mean MW of 1.4 .times. 105. The synthesis of this RNA was suppressed, but apparently not completely so, by inhibition of cellular DNA synthesis to more than 95% by addition of cytosine arabinoside. The synthesis of other RNA species remained unaffected by this treatment. Poly(A)-containing mRNA, representing a 20% contaminant of the electrophoretically purified histone mRNA, was removed by poly(U)-Sepharose chromatography. Histone mRNA thus purified was hybridized to DNA fractions enriched for histone genes of the sea urchin Psammechinus miliaris by actinomycin/CsCl gradient centrifugation. The mRNA was eluted from the hybrids and challenged with mouse DNA in vast DNA excess conditions. A c0t1/2 [initial denatured DNA concentration] of 360 mol s l-1 was obtained from the RNA trace curve, suggesting a 10-20-fold reiteration of the histone genes in the haploid genome. Thus histone genes in mouse are much less highly reiterated than in sea urchins, but nevertheless are present in considerable excess over the number theoretically necessary for histone synthesis during the S phase of the cell cycle.