Measurement of the linking number change in transcribing chromatin

The in vivo-initiated, transcribing simian virus 40 (SV40) minichromosome was analyzed to determine its DNA linking number change, i.e. the difference between the linking number of the minichromosomal DNA and that of relaxed bare DNA. As part of this measurement, the linking number change due to the in vivo-initiated RNA polymerase II was determined, the first time a value for this quantity has been reported. The topological contribution of the polymerase was combined with values determined for constrained and non-constrained linking number contributions from the native transcription complex chromatin to yield the linking number change for the complex. The linking number change of the native non-transcribed SV40 minichromosome was independently determined and was found to be virtually the same as that for the chromatin of the transcription complex. This indicates that there is little difference between the two structures. The plausibility of several current models for the contribution of chromatin structure to transcription regulation is discussed in light of this finding.