In vivoincorporation ofDrosophilaH2a histone into mammalian chromatin | Zendy

Raymond Reeves | Zendy; Cornelia M. Gorman | Zendy; Bruce H. Howard | Zendy

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In vivoincorporation ofDrosophilaH2a histone into mammalian chromatin

Author(s) -

Raymond Reeves,

Cornelia M. Gorman,

Bruce H. Howard

Publication year - 1983

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/11.9.2681

Subject(s) - biology , chromatin , histone h2a , histone , gene , schneider 2 cells , polyadenylation , drosophila melanogaster , nucleosome , microbiology and biotechnology , genetics , rna interference , gene expression , rna

Hybrid prokaryotic/eukaryotic expression vectors have been used to introduce Drosophila histone genes into CV-1 African green monkey tissue culture cells. Transfection of CV-1 cells with Drosophila genes under the control of insect DNA promoter sequences results in low level expression of histone genes. On the other hand, when the Drosophila H2a gene is juxtaposed downstream from the long terminal repeat sequence of Rous sarcoma virus (RSV) expression of the insect gene is considerably more efficient; both 3' polyadenylated insect histone messenger RNA and putative Drosophila H2a histone protein can be readily detected in the transduced cells. Using this RSV/H2a vector, we have been able to demonstrate the presence of Drosophila H2a histone in monomer nucleosome preparations isolated from transfected CV-1 cells. These results suggest the feasibility of 'remodeling' cellular chromatin in vivo in precisely defined ways. The techniques described may be generally applicable to other genes coding for chromosomal proteins.

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