PLAMseq enables the proteo-genomic characterization of chromatin-associated proteins and protein interactions in a single experimental workflow.

Gonzalez-Vinceiro, L.; Espejo-Serrano, C.; Soler-Oliva, M. E.; Mateos-Martin, M. L.; Rico, D.; Gonzalez-Aguilera, C.; Gonzalez Prieto, R.

doi:10.1101/2025.04.27.650851

bioRxiv · 10.1101/2025.04.27.650851

PLAMseq enables the proteo-genomic characterization of chromatin-associated proteins and protein interactions in a single experimental workflow.

Gonzalez-Vinceiro, L.·Espejo-Serrano, C.·Soler-Oliva, M. E.·Mateos-Martin, M. L.·Rico, D.·Gonzalez-Aguilera, C.·Gonzalez Prieto, R.

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Abstract

Chromatin Immunoprecipitation (ChIP) and Co-Immunoprecipitation (CoIP) assays are common approaches to characterize the genomic localization and protein interactors, respectively, for a protein of interest. However, these approaches require the use of specific antibodies, which often face sensitivity and specificity issues. Based on TurboID, we developed PLAMseq (Proximity Labelled Affinity-purified Mass spectrometry plus sequencing), which enables, in the same workflow, to identify the genomic loci and the interacting proteome of a protein of interest. Moreover, PLAMseq can also be applied to specifically map protein interactions and ubiquitin(-like) modified proteins. We validated PLAMseq with two well characterized proteins, RNA polymerase II and CTCF, with excellent robustness and reproducibility. Next, we applied PLAMseq to characterize Histone H1 SUMOylation, which study has remained elusive due to the lack of specific reagents, and found that SETDB1 binds to SUMOylated histone H1.2 and H1.4 which also colocalize with H3K9me3 at repetitive regions of the genome.

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Gonzalez-Vinceiro, L., Espejo-Serrano, C., Soler-Oliva, M. E., Mateos-Martin, M. L., Rico, D., Gonzalez-Aguilera, C., Gonzalez Prieto, R.. 2025-04-27. PLAMseq enables the proteo-genomic characterization of chromatin-associated proteins and protein interactions in a single experimental workflow.. https://doi.org/10.1101/2025.04.27.650851

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