Abstract
How evolutionary changes at enhancers affect the transcription of target genes remains an important open question. Previous comparative studies of gene expression have largely measured the abundance of messenger RNA, which is affected by post-transcriptional regulatory processes, hence limiting inferences about the mechanisms underlying expression differences. Here, we directly measured nascent transcription in primate species, allowing us to separate transcription from post-transcriptional regulation. We used precision run-on and sequencing to map RNA polymerases in resting and activated CD4+ T cells in multiple human, chimpanzee and rhesus macaque individuals, with rodents as outgroups. We observed general conservation in coding and non-coding transcription, punctuated by numerous differences between species, particularly at distal enhancers and non-coding RNAs. Genes regulated by larger numbers of enhancers are more frequently transcribed at evolutionarily stable levels, despite reduced conservation at individual enhancers. Adaptive nucleotide substitutions are associated with lineage-specific transcription and at one locus, SGPP2, we predict and experimentally validate that multiple substitutions contribute to human-specific transcription. Collectively, our findings suggest a pervasive role for evolutionary compensation across ensembles of enhancers that jointly regulate target genes.
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Acknowledgements
We thank M. Jin for assistance in establishing the magnetic separation of CD4+ T cells, J. Rogers for help establishing contacts with primate centres, L. Core, H. Kwak, N. Fuda and I. Jonkers for assistance troubleshooting the PRO-Seq library prep, and A. Wetterau for preparing nuclei for mouse and rat CD4+ T cells. This work was supported by generous seed grants from the Cornell University Center for Vertebrate Genomics, Center for Comparative and Population Genetics, National Human Genome Research Institute grant HG009309 to C.G.D., National Heart, Lung, and Blood Institute grant UHL129958A to C.G.D. and J.T.L., National Institute of General Medical Sciences grant GM102192 to A.S., National Human Genome Research Institute grant HG0070707 to A.S. and J.T.L., National Institute of Diabetes and Digestive and Kidney Diseases DK058110 to W.L.K. and Cancer Prevention and Research Institute of Texas RP160319 to W.L.K. The content is solely the responsibility of the authors and does not necessarily represent the official views of the US National Institutes of Health. Finally, we would like to thank the anonymous human and non-human primate donors who gave blood in support of this study.
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L.A.C., B.A.M., C.G.D., E.J.R. and E.D.T.W. performed the CD4+ T cell extraction, validation and PRO-Seq experiments. C.G.D., Z.W., T.C., A.L.M., L.A.C. and N.D. analysed the data. C.G.D., A.S., J.T.L., W.L.K. and S.A.C. supervised the data collection and analysis. C.G.D. and A.S. wrote the paper with input from the other authors.
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Danko, C.G., Choate, L.A., Marks, B.A. et al. Dynamic evolution of regulatory element ensembles in primate CD4+ T cells. Nat Ecol Evol 2, 537–548 (2018). https://doi.org/10.1038/s41559-017-0447-5
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DOI: https://doi.org/10.1038/s41559-017-0447-5
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