RNA polymerase II pausing temporally coordinates cell cycle progression and erythroid differentiation

Danya J Martell; Hope E Merens; Alexis Caulier; Claudia Fiorini; Jacob C Ulirsch; Robert Ietswaart; Karine Choquet; Giovanna Graziadei; Valentina Brancaleoni; Maria Domenica Cappellini; Caroline Scott; Nigel Roberts; Melanie Proven; Noémi B A Roy; Christian Babbs; Douglas R Higgs; Vijay G Sankaran; L Stirling Churchman

doi:10.1016/j.devcel.2023.07.018

RNA polymerase II pausing temporally coordinates cell cycle progression and erythroid differentiation

Dev Cell. 2023 Oct 23;58(20):2112-2127.e4. doi: 10.1016/j.devcel.2023.07.018. Epub 2023 Aug 15.

Authors

Danya J Martell¹, Hope E Merens², Alexis Caulier³, Claudia Fiorini³, Jacob C Ulirsch³, Robert Ietswaart², Karine Choquet², Giovanna Graziadei⁴, Valentina Brancaleoni⁴, Maria Domenica Cappellini⁴, Caroline Scott⁵, Nigel Roberts⁶, Melanie Proven⁷, Noémi B A Roy⁸, Christian Babbs⁵, Douglas R Higgs⁵, Vijay G Sankaran⁹, L Stirling Churchman¹⁰

Affiliations

¹ Department of Genetics, Harvard University, Boston, MA, USA; Division of Hematology/Oncology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA; Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, USA; Broad Institute of MIT and Harvard, Cambridge, MA, USA.
² Department of Genetics, Harvard University, Boston, MA, USA.
³ Division of Hematology/Oncology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA; Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, USA; Broad Institute of MIT and Harvard, Cambridge, MA, USA.
⁴ Department of Clinical Sciences and Community, University of Milan, IRCCS Ca'Granda Foundation Maggiore Policlinico Hospital, Milan, Italy.
⁵ MRC Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, UK.
⁶ MRC Molecular Haematology Unit, MRC Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, UK.
⁷ Oxford Genetics Laboratories, Oxford University Hospitals NHS Foundation Trust, Oxford, UK.
⁸ MRC Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, UK; NIHR Oxford Biomedical Research Centre and BRC/NHS Translational Molecular Diagnostics Centre, John Radcliffe Hospital, Oxford, UK; Department of Haematology, Oxford University Hospitals NHS Foundation Trust, Oxford, UK.
⁹ Division of Hematology/Oncology, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA; Department of Pediatric Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, USA; Broad Institute of MIT and Harvard, Cambridge, MA, USA. Electronic address: [email protected].
¹⁰ Department of Genetics, Harvard University, Boston, MA, USA. Electronic address: [email protected].

Abstract

Controlled release of promoter-proximal paused RNA polymerase II (RNA Pol II) is crucial for gene regulation. However, studying RNA Pol II pausing is challenging, as pause-release factors are almost all essential. In this study, we identified heterozygous loss-of-function mutations in SUPT5H, which encodes SPT5, in individuals with β-thalassemia. During erythropoiesis in healthy human cells, cell cycle genes were highly paused as cells transition from progenitors to precursors. When the pathogenic mutations were recapitulated by SUPT5H editing, RNA Pol II pause release was globally disrupted, and as cells began transitioning from progenitors to precursors, differentiation was delayed, accompanied by a transient lag in erythroid-specific gene expression and cell cycle kinetics. Despite this delay, cells terminally differentiate, and cell cycle phase distributions normalize. Therefore, hindering pause release perturbs proliferation and differentiation dynamics at a key transition during erythropoiesis, identifying a role for RNA Pol II pausing in temporally coordinating the cell cycle and erythroid differentiation.

Keywords: RNA Pol II pausing; SPT5; SUPT5H; cell cycle; differentiation; erythropoiesis; hemoglobin; human genetic variation; transcriptional regulation.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Cell Cycle
Cell Differentiation
Gene Expression Regulation*
Humans
Nuclear Proteins / metabolism
RNA Polymerase II* / genetics
RNA Polymerase II* / metabolism
Transcription, Genetic
Transcriptional Elongation Factors / genetics

Substances

RNA Polymerase II
SUPT5H protein, human
Nuclear Proteins
Transcriptional Elongation Factors

Abstract

Publication types

MeSH terms

Substances

Grants and funding