Nucleic Acids Research, Vol 24, Issue 7 1212-1219, Copyright © 1996 by Oxford University Press
DA Jackson, J Bartlett and PR Cook
DNA sequences attaching loops of nuclear and mitochondrial DNA to
underlying structures in HeLa cells have been cloned and 106 representative
clones sequenced; 10 clones containing random genomic fragments served as
controls. As chromatin is prone to rearrangement, care was taken to isolate
sequences using 'physiological' conditions that did not create additional
attachments. Comparison (by Southern blotting) of the concentration of each
cloned sequence in 'total' and 'attached' fractions of DNA showed that most
clones did contain attached sequences, but even highly-attached sequences
were not attached in all cells in the population. Results demonstrated that
28% of clones were derived from three specific parts of the mitochondrial
genome and 22% from different parts of the alu repeat. In addition, 41% of
clones contained unique nuclear sequences; these contained no more of the
motifs found attached to nuclear scaffolds or matrices (ie SARs or MARs)
than would be expected from their base composition. No other attachment
motif(s) could be identified by sequence analysis. However, Northern
blotting showed that all the mitochondrial clones and 76% of clones
containing unique sequences were transcribed; the degree of attachment
correlated with transcriptional activity. These results are consistent with
transcription being responsible for ever-changing attachments in both
nuclei and mitochondria.
ARTICLES
Sequences attaching loops of nuclear and mitochondrial DNA to underlying structures in human cells: the role of transcription units
CRC Nuclear Structure and Function Research Group, Sir William Dunn School of Pathology, University of Oxford, UK.
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