Nucleic Acids Research, 1986, Vol. 14, No. 22 8845-8862
© 1986
Articles |
A chimeric mouse histone H4 gene containing either an intron or poly(A) addition signal behaves like a basal histone
Laboratory of Biochemistry, National Cancer Institute, National Institutes of Health Bethesda, MD 20892, USA
Received August 28, 1986. Revised October 20, 1986. Accepted October 20, 1986.
We have modified the basic structure of the mouse H4 histone gene by introducing, in one case, the IVS-II of the human beta globin gene in the middle of the H4 coding region and, in the second case, the poly(A) addition signal from either the chicken vimentin gene or the alpha globin gene, displacing the hairpin loop structure in the 3 per 10 thousand direction. Constructs were placed into the vector, PSV2gpt, and stably transformed into L cells. Pools of 100–500 independent transformants were analyzed for H4 expression. Even though the intron is processed correctly, the growth regulated expression of the modified gene is lost and the gene is now expressed at a constant basal level. Furthermore, unprocessed transcripts accumulate in the nucleus of Go cells when compared to exponentially growing cultures. Polyadenylated Hi RNA is correctly processed but expressed at reduced levels (30 fold) in a constitutive manner, independent of the growth state of the cell. The altered expression of these chimeric H4 gene3 compared to the endogenous copy or the transfected wild type gene suggests a structural model to explain the cell cycle independent expression of the basal histones.
*Present address: Université de Lausanne, Institut de Biologie Animale, Bâltiment de Biologie, CH-1015 Lausanne, Switzerland