RESEARCH
Recent Works
Explore a selection of my academic papers and significant contributions to the field of biology, focusing on DNA replication through experimental research with yeast and light microscopy techniques.
Stabilization of expandable DNA repeats by the replication factor Mcm10 promotes cell viability
Chiara Masnovo, Zohar Paleiov, Daniel Dovrat, Laurel K. Baxter,Sofia Movafaghi, Amir Aharoni & Sergei M. Mirkin
This study demonstrates that the replication factor Mcm10 plays a crucial role in preventing instability of expanded GAA trinucleotide repeats (associated with Friedreich's ataxia) by maintaining proper interaction with the CMG helicase during DNA replication through these repetitive sequences. It was found that Mcm10 deficiency causes replication fork stalling at GAA repeats, leading to cell death when the repeats are located on essential chromosome regions, though cells can survive through Rad9 checkpoint-mediated repair that simultaneously promotes repeat expansions.
Transcription-replication coordination revealed in single live cells
Ioannis Tsirkas, Daniel Dovrat, Manikandan Thangaraj, Ineke Brouwer, Amit Cohen, Zohar Paleiov, Michael M. Meijler, Tineke Lenstra & Amir Aharoni
This study utilized a live-cell imaging approach to simultaneously monitor DNA replication fork progression and transcription dynamics in single S.cerevisiae cells, revealing that transcription is partially repressed in a wave that travels 27-30 kb ahead of the advancing replication fork. While this pre-emptive transcriptional repression allows smooth fork progression through highly transcribed genes under normal conditions, mutations that impair replication-transcription coordination lead to conflicts causing fork stalling and reduced transcription, with cells showing trade-offs between maintaining replication speed versus transcription levels.
Enhanced fluorescent imaging of proteins in live yeast cells using fluorescently labeled scFv
Ioannis Tsirkas, Tomer Zur, Daniel Dovrat, Zohar Paleiov, Lior Ravkaie & Amir Aharoni
This protocol paper presents a method for fluorescently labeling HA-tagged proteins in live yeast cells using the single-chain antibody (scFv) 2E2 fused to various fluorescent proteins, enabling enhanced visualization of proteins across different cellular compartments and expression levels. The approach provides a collection of plasmids containing 2E2-FP fusions and detailed steps for implementation of this labeling system to examine protein function, expression, and localization through live-cell microscopy in S.cerevisiae.