Molecular Cell Engineering Laboratory

Kanemaki Group

Studying DNA replication by use of an original degron-based technology

Faculty

KANEMAKI, Masato
Professor

mkanemak@nig.ac.jp

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YAMAGUCHI, Kosuke
Assistant Professor

kyamaguchi@nig.ac.jp

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 Lab HP

Research Summary

Our laboratory pioneered the development of the auxin-inducible degron (AID) technology, which enables the rapid degradation of degron-tagged proteins of interest upon the addition of the plant hormone auxin. We recently improved this system and established AID2, which allows controlled degradation of target proteins not only in yeast and mammalian cell lines, but also in mice. By applying AID2, we aim to elucidate the mechanisms of DNA replication in human cells and mice.

Replication initiation zone mapping using AID2. (A) Our laboratory developed the Ligase-depletion Okazaki fragment sequencing (LD-OK-seq) method to detect replication initiation zones genome-wide. When DNA ligases in cells are degraded using AID2, Okazaki fragments synthesized at replication forks accumulate. Sequencing these fragments enables the determination of replication fork directionality and the identification of replication initiation zones. (B) An example of LD-OK-seq data. Replication fork directionality and replication initiation zones can be detected with high precision.

Selected Publications

  • Zhu, X., Bektash, A., Hatoyama, Y., Muramatsu, S., Isobe, S.-Y., Obuse, C., Toyoda, A., Daigaku, Y., Chen, C.-L., and Kanemaki, M.T. (2025). Regulated TRESLIN-MTBP loading governs initiation zones and replication timing in human DNA replication. Nat. Commun. 16, 10069. https://doi.org/10.1038/s41467-025-66278-7.
  • Islam, M., Negishi, T., Kitamoto, N., Hatoyama, Y., Gamo, K., Hayashi, K., and Kanemaki, M.T. (2025). A single-chain antibody-based AID2 system for conditional degradation of GFP-tagged and untagged proteins. J. Cell Sci. 138. https://doi.org/10.1242/jcs.263961.
  • Hatoyama, Y., Islam, M., Bond, A.G., Hayashi, K., Ciulli, A., and Kanemaki, M.T. (2024). Combination of AID2 and BromoTag expands the utility of degron-based protein knockdowns. EMBO Rep. 25, 4062–4077. https://doi.org/10.1038/s44319-024-00224-4.
  • Saito, Y., Santosa, V., Ishiguro, K., and Kanemaki, M.T. (2022). MCMBP promotes the assembly of the MCM2–7 hetero-hexamer to ensure robust DNA replication in human cells. eLife 11, e77393. https://doi.org/10.7554/elife.77393.