Contribution of Growing RNA Molecules to the Nuclear Transcripts Foci Observed by FISH

Abstract

The focal distributions of RNAs observed using fluorescencein situhybridization (FISH) in the cell nucleus may correspond either to RNAs in the course of transcription or essentially to accumulation of full-length transcripts at the sites of transcription. To determine to what extent nuclear transcript foci represent growing RNA molecules, uninterrupted hsp70 transcripts were detected by FISH in heat-shocked human fibroblasts using two probes specific for the 5′ or for the 3′ end of the transcripts. By comparing the size of the signals obtained with each probe, we show that transcript foci mainly represent accumulations of full-length transcripts at their site of transcription. A major contribution of RNAs in the course of transcription to the transcript foci is observed only during the first minutes of gene induction. These observations suggest the existence of a rate-limiting step in the release of newly synthesized transcripts from their site of transcription, which is independent from the step of intron excision.