Regulated Expression of a Chimeric Histone Gene Introduced into Mouse Fibroblasts

Alterman, R B; Sprecher, Cindy A.; Graves, Reed A.; Marzluff, William F.; Skoultchi, Arthur I.

doi:10.1128/mcb.5.9.2316-2324.1985

Molecular and Cellular Biology

1985

DOI: 10.1128/mcb.5.9.2316-2324.1985

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Regulated Expression of a Chimeric Histone Gene Introduced into Mouse Fibroblasts

R B Alterman¹,

Cindy A. Sprecher²,

Reed A. Graves³

et al.

Abstract: The regulated expression of a mouse histone gene was studied by DNA-mediated gene transfer. A chimeric H3 histone gene was constructed by fusing the 5' and 3' portions of two different mouse H3 histone genes. Transfection of the chimeric gene into mouse fibroblasts resulted in the production of chimeric mRNA at levels nearly equal to that of the total endogenous H3 histone mRNAs. Most chimeric RNA transcripts had correct 5' and 3' termini, and the chimeric mRNA was translated into an H3.1 protein that accumula… Show more

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1986

1988

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Cited by 2 publications

References 25 publications

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Histone mRNA Degradation In Vivo: the First Detectable Step Occurs at or near the 3′ Terminus

Ross¹,

Peltz²,

Kobs³

et al. 1986

Molecular and Cellular Biology

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The first detectable step in the degradation of human H4 histone mRNA occurs at the 3' terminus in a cell-free mRNA decay system (J. Ross and G. Kobs, J. Mol. Biol. 188:579-593, 1986). Most or all of the remainder of the mRNA is then degraded in a 3'-to-5' direction. The experiments described here were designed to determine whether a similar degradation pathway is followed in whole cells. Two sets of short-lived histone mRNA decay products were detected in logarithmically growing erythroleukemia (K562) cells. These products, designated the -5 and -12 RNAs, were generated by the loss of approximately 4 to 6 and 11 to 13 nucleotides, respectively, from the 3' terminus of histone mRNA. The same decay products were observed after a brief incubation in vitro. They were in low abundance or absent from cells that were not degrading histone mRNA. In contrast, they were readily detectable in cells that degraded the mRNA at an accelerated rate, i.e., in cells cultured with a DNA synthesis inhibitor, either cytosine arabinoside or hydroxyurea. During the initial stages of the decay process, as the 3' terminus of the mRNA was being degraded, the 5'-terminal region remained intact. These results indicate that the first detectable step in human H4 histone mRNA decay occurs at the 3' terminus and that degradation proceeds 3' to 5', both in cells and in cell-free reactions.

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Histone mRNA Degradation In Vivo: the First Detectable Step Occurs at or near the 3′ Terminus

Ross¹,

Peltz²,

Kobs³

et al. 1986

Molecular and Cellular Biology

View full text Add to dashboard Cite

show abstract

Differential Expression of Individual Members of the Histone Multigene Family Due to Sequences in the 5′ and 3′ Regions of the Genes

Levine

Liu

Marzluff

et al. 1988

Molecular and Cellular Biology

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Histone proteins are encoded by a multigene family. The H3.2(614) and H2a(614) genes are present as single copies which are expressed at high levels, accounting for 30 to 40% of the H3 and H2a mRNAs, respectively, in different types of mouse cells. The other genes which have been isolated each contribute only a very small amount to the total type-specific mRNA pool. We demonstrate here that the differences in the level of expression of these genes are partly due to differences in their transcription rates. To investigate the sequences responsible for these differences in expression among the members of each family, we carried out DNA-mediated gene transfer experiments with both intact and chimeric histone genes. The 5' region of a highly expressed gene [H3.2(614) or H2a(614)] was attached to the 3' region of a histone gene which was expressed at low levels (H3-221 or H2a-291) and vice versa. The results show that sequences in both the 5' and 3' regions of the H3.2(614) and H2a(614) genes contribute to their high level of mRNA production by two independent mechanisms. The effect of the 3' sequences on mRNA accumulation has been narrowed to a 65-base-pair region including the 3'-terminal palindrome and downstream signal implicated in mRNA processing.

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Regulated Expression of a Chimeric Histone Gene Introduced into Mouse Fibroblasts

Cited by 2 publications

References 25 publications

Histone mRNA Degradation In Vivo: the First Detectable Step Occurs at or near the 3′ Terminus

Histone mRNA Degradation In Vivo: the First Detectable Step Occurs at or near the 3′ Terminus

Differential Expression of Individual Members of the Histone Multigene Family Due to Sequences in the 5′ and 3′ Regions of the Genes

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