Transcriptional repression by methylation: cooperativity between a CpG cluster in the promoter and remote CpG-rich regions.

Abstract:

:Cytosine methylation of binding sites for transcription factors is a straightforward mechanism to prevent transcription, while data on an indirect mechanism, by methylation outside of the factor binding sites, are still scarce. We have studied the latter effect using a model promoter construct. For this, a 69 bp G + C rich DNA segment with a cluster of 14 CpG sites was inserted between upstream lexA sites and the TATA box. Transcription was measured in transient transfection assays with lexA-VP16 as an activating factor. When the entire plasmid was methylated at all CpGs before transfection, transcription was blocked (to 3% residual activity), whereas transcription was only mildly inhibited (to 60%) by methylation of a control plasmid that lacked the 69 bp CpG cluster. However, the effect could not simply be attributed to methylation of the CpG cluster: neither a methylated CpG cluster in an otherwise methylation-free reporter gene plasmid, nor the methylated plasmid with an unmethylated CpG cluster, inhibited transcription considerably (69% and 44% remaining activity, respectively). The data presented here suggest that a minimal length of methylated DNA in the promoter is required for repression, and imply that concomitant methylation of CpGs in the promoter region and in remote sequences can cooperatively block transcription, without the need to methylate any binding sites for transcription factors. We also note that the cooperation for a negative effect described here bears an analogy to transcriptional activation, where a promoter often cooperates with a remote enhancer.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Hug M,Silke J,Georgiev O,Rusconi S,Schaffner W,Matsuo K

doi

10.1016/0014-5793(95)01521-3

subject

Has Abstract

pub_date

1996-02-05 00:00:00

pages

251-4

issue

3

eissn

0014-5793

issn

1873-3468

pii

0014-5793(95)01521-3

journal_volume

379

pub_type

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