Honors & Awards
EMBO postdoctoral fellowship, European Molecular Biology Organization (2023-2024)
School of Medicine Dean's fellowship, Stanford University (2023)
GENE Mentors fellowship, EUR GENE, Université Paris Cité (2022)
DOC4 fellowship, Fondation ARC pour la recherche sur le cancer (2020-2021)
Bourse de thèse (PhD fellowship), French Ministery for Education and Research (2017-2020)
International Masters Mobility Scholarship (MIEM) program, Université Paris Sorbonne Cité (2016)
PhD, Université Paris Cité, Genetics (2021)
MSc, Université Denis Diderot - Paris VII, Genetics (2017)
MSc, Sapienza, University of Rome, Genetics and molecular biology (2017)
BSc, Sapienza, University of Rome, Biology (2015)
Lars Steinmetz, Postdoctoral Faculty Sponsor
Sen1 is a key regulator of transcription-driven conflicts.
Cellular homeostasis requires the coordination of several machineries concurrently engaged in the DNA. Wide-spread transcription can interfere with other processes, and transcription-replication conflicts (TRCs) threaten genome stability. The conserved Sen1 helicase not only terminates non-coding transcription but also interacts with the replisome and reportedly resolves genotoxic R-loops. Sen1 prevents genomic instability, but how this relates to its molecular functions remains unclear. We generated high-resolution, genome-wide maps of transcription-dependent conflicts and R-loops using a Sen1 mutant that has lost interaction with the replisome but is termination proficient. We show that, under physiological conditions, Sen1 removes RNA polymerase II at TRCs within genes and the rDNA and at sites of transcription-transcription conflicts, thus qualifying as a "key regulator of conflicts." We demonstrate that genomic stability is affected by Sen1 mutation only when in addition to its role at the replisome, the termination of non-coding transcription or R-loop removal are additionally compromised.
View details for DOI 10.1016/j.molcel.2022.06.021
View details for PubMedID 35839782
DNA supercoiling restricts the transcriptional bursting of neighboring eukaryotic genes.
2023; 83 (10): 1573-1587.e8
DNA supercoiling has emerged as a major contributor to gene regulation in bacteria, but how DNA supercoiling impacts transcription dynamics in eukaryotes is unclear. Here, using single-molecule dual-color nascent transcription imaging in budding yeast, we show that transcriptional bursting of divergent and tandem GAL genes is coupled. Temporal coupling of neighboring genes requires rapid release of DNA supercoils by topoisomerases. When DNA supercoils accumulate, transcription of one gene inhibits transcription at its adjacent genes. Transcription inhibition of the GAL genes results from destabilized binding of the transcription factor Gal4. Moreover, wild-type yeast minimizes supercoiling-mediated inhibition by maintaining sufficient levels of topoisomerases. Overall, we discover fundamental differences in transcriptional control by DNA supercoiling between bacteria and yeast and show that rapid supercoiling release in eukaryotes ensures proper gene expression of neighboring genes.
View details for DOI 10.1016/j.molcel.2023.04.015
View details for PubMedID 37207624
An integrated model for termination of RNA polymerase III transcription.
2022; 8 (28): eabm9875
RNA polymerase III (RNAPIII) synthesizes essential and abundant noncoding RNAs such as transfer RNAs. Controlling RNAPIII span of activity by accurate and efficient termination is a challenging necessity to ensure robust gene expression and to prevent conflicts with other DNA-associated machineries. The mechanism of RNAPIII termination is believed to be simpler than that of other eukaryotic RNA polymerases, solely relying on the recognition of a T-tract in the nontemplate strand. Here, we combine high-resolution genome-wide analyses and in vitro transcription termination assays to revisit the mechanism of RNAPIII transcription termination in budding yeast. We show that T-tracts are necessary but not always sufficient for termination and that secondary structures of the nascent RNAs are important auxiliary cis-acting elements. Moreover, we show that the helicase Sen1 plays a key role in a fail-safe termination pathway. Our results provide a comprehensive model illustrating how multiple mechanisms cooperate to ensure efficient RNAPIII transcription termination.
View details for DOI 10.1126/sciadv.abm9875
View details for PubMedID 35857496
Sen1 Is Recruited to Replication Forks via Ctf4 and Mrc1 and Promotes Genome Stability
2020; 30 (7): 2094-2105
DNA replication and RNA transcription compete for the same substrate during S phase. Cells have evolved several mechanisms to minimize such conflicts. Here, we identify the mechanism by which the transcription termination helicase Sen1 associates with replisomes. We show that the N terminus of Sen1 is both sufficient and necessary for replisome association and that it binds to the replisome via the components Ctf4 and Mrc1. We generated a separation of function mutant, sen1-3, which abolishes replisome binding without affecting transcription termination. We observe that the sen1-3 mutants show increased genome instability and recombination levels. Moreover, sen1-3 is synthetically defective with mutations in genes involved in RNA metabolism and the S phase checkpoint. RNH1 overexpression suppresses defects in the former, but not the latter. These findings illustrate how Sen1 plays a key function at replication forks during DNA replication to promote fork progression and chromosome stability.
View details for DOI 10.1016/j.celrep.2020.01.087
View details for Web of Science ID 000514824500005
View details for PubMedID 32075754
View details for PubMedCentralID PMC7034062