Generation of recombinant constructs pECFP-C3-USP1-domain-1 and pECFP-C3-USP1-domain-2 as key tools for studying the domain organization of USP1 and its interaction with the oncoprotein Bcr-Abl

  • A. S. Matvieienko Institute of Molecular Biology and Genetics of NAS of Ukraine, Ukraine, 03143, Kyiv, Akademika Zabolotnogo str., 150; Kyiv National University of Technologies and Design, Ukraine, 01011, Kyiv, Mala Shiyanovska str., 2
  • S. V. Antonenko Institute of Molecular Biology and Genetics of NAS of Ukraine, Ukraine, 03143, Kyiv, Akademika Zabolotnogo str., 150 https://orcid.org/0000-0003-0212-3779
  • D. S. Gur’yanov Institute of Molecular Biology and Genetics of NAS of Ukraine, Ukraine, 03143, Kyiv, Akademika Zabolotnogo str., 150 https://orcid.org/0000-0003-0594-2286
  • G. D. Telegeev Institute of Molecular Biology and Genetics of NAS of Ukraine, Ukraine, 03143, Kyiv, Akademika Zabolotnogo str., 150 https://orcid.org/0000-0003-0270-4397
DOI: https://doi.org/10.7124/FEEO.v36.1715
Keywords: chronic myeloid leukemia (CML), Bcr-Abl oncoprotein, ubiquitin-specific proteases (USPs), USP1, USP1 domains

Abstract

Aim. The aim of this study is to generate recombinant constructs pECFP-C3-USP1-domain-1 and pECFP-C3-USP1-domain-2, essential for investigating the domain organization of the USP1 protein, determining their role in USP1 function, and analyzing their interaction with the oncoprotein Bcr-Abl. Methods. Standard molecular cloning techniques were employed, including polymerase chain reaction (PCR), ligation, restriction enzyme digestion, as well as DNA extraction and purification methods. Results. The genetic constructs pECFP-C3-USP1-domain-1 and pECFP-C3-USP1-domain-2, encoding key domains of the USP1 protein, were successfully designed and obtained. Conclusions. The generated genetic constructs pECFP-C3-USP1-domain-1 and pECFP-C3-USP1-domain-2 will be utilized to study the role of these domains in USP1 function, its subcellular localization, and the formation of the Bcr-Abl/USP1 protein complex. These findings will contribute to a better understanding of the role of USP1 in cellular processes and facilitate the development of novel therapeutic approaches for the treatment of chronic myeloid leukemia (CML).

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