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DNA conformation and chromatin organization of a d(CA/GT)30 sequence cloned in SV40 minichromosomes.
Author(s) -
RodriguezCampos A.,
Ellison M.J.,
PérezGrau L.,
Azorin F.
Publication year - 1986
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1986.tb04417.x
Subject(s) - biology , chromatin , minichromosome , dna , sequence (biology) , genetics , microbiology and biotechnology , dna sequencing
The alternating DNA sequence d(CA/GT)n is known to adopt the left‐handed Z‐form in negatively supercoiled DNA in vitro. This element represents a significant fraction of the highly repeated DNA sequences found in eukaryotic genomes. We have cloned an alternating d(CA/GT)30 sequence into SV40 minichromosomes at the unique HpaII restriction site which occurs in the transcriptional leader region of the viral late genes. By comparing the linkage differences of topoisomers obtained from viral DNA with or without the d(CA/GT)30 insert, at various stages of the lytic cycle, we conclude that this sequence does not predominate in the Z‐form in vivo. Furthermore, we find that the d(CA/GT)30 sequence is packaged into nucleosomal core particles and the region of the minichromosomes which contain the d(CA/GT)30 sequence is organized as nucleosomes.