Deoxyribonuclease I sensitivity of the T-DNA ipt gene is associated with gene expression

Abstract

We have analyzed the chromatin structure of the T-DNA isopentenyl transferase gene, ipt, in four Nicotiana tabacum crown gall tumor lines. These four transformed lines contain identical T-DNA inserts and are derivatives of a single clone that did not exhibit any tumorous properties and contained a highly methylated, nonexpressed copy of T-DNA. One of the derivatives also does not exhibit tumorous properties, and the T-DNA of this line is not expressed. The other three lines have reverted to tumorous growth either spontaneously or after treatment with the inhibitor of DNA methylation, 5-azacytidine. Concomitant with this reversion to tumorous growth, expression of the ipt gene of these lines has reinitiated. In the lines that express the ipt gene, the chromatin structure of this gene exists in a conformation that is more accessible to DNase I than in the line in which this gene is not expressed. The level of ipt expression and DNase I sensitivity was independent of the process by which the transformed cell lines reverted to tumorous growth. The relationship of chromatin structure to gene expression and DNA methylation in these lines is discussed.