A number of transcriptional and chromatin regulatory proteins have been reported to be recruited to chromatin by specific regulatory RNAs. Whether RNA has a more general role in the interaction of proteins with chromatin is unknown. We used proteomics methods to determine the global impact of nascent RNA on chromatin in embryonic stem cells. Surprisingly, we found that nascent RNA primarily antagonises the interaction of chromatin modifiers and transcriptional regulators with chromatin. Transcriptional inhibition and RNA degradation induced recruitment of a set of transcriptional regulators, chromatin modifiers, nucleosome remodelers, and regulators of higher-order structure. RNA directly bound to factors including BAF, NuRD, EHMT1 and INO80 and inhibited their interaction with nucleosomes. In the case of the transcriptional elongation factor P-TEFb, direct binding to pre-mRNA released it from the chromatin-associated 7SK ribonucleoprotein complex. We propose that through these mechanisms, nascent RNA provides a direct link between transcriptional output and chromatin state.