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Nucleic Acids Research, 1989, Vol. 17, No. 24 10179-10190
© 1989

Articles

Cytosine methylation in CTF and Spl recognition sites of an HSV tk promoter: effects on transcription in vivo and on factor binding in vitro

Jean Ben-Hattar1,2, Peter Beard1 and Josef Jiricny2

1Swiss Institute for Experimental Cancer Research CH-1066 Epalinges s/Lausanne 2Friedrich Miescher-Institut CH-4002 Basel, Switzerland

Received October 23, 1989. Accepted November 16, 1989.

We methylated specific cytosinc residues within or immediately around the CTF and Spl binding sites of the Herpes simplex virus thymidine kinase promoter. The efficiency of transcription in vivo was reduced at least 50-fold compared with transcription from the unmethylated promoter. However, methylation within the CTF recognition site had no effect on the affinity of CTF for this site in vitro. Mthylation of the Spl site resulted in only a small decrease in the affinity of this factor for its recognition site. In vivo studies showed that the same gene inserted in different vector DNAs was regulated differently by methylation in the promoter. These results show that cytosine methylation can inhibit transcription by a mechanism other than directly blocking the binding of transcription factors.


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