Osadchiy Igor, Umnova Anastasia, Pokholkova Galina V, Golovnin Anton, Gvozdev Vladimir A, Zhimulev Igor F, Georgiev Pavel, Maksimenko Oksana
Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Institute of Gene Biology, Russian Academy of Sciences, 34/5 Vavilov St., Moscow, 119334, Russia.
Department of the Control of Genetic Processes, Institute of Gene Biology, Russian Academy of Sciences, 34/5 Vavilov St., Moscow, 119334, Russia.
Epigenetics Chromatin. 2025 Apr 16;18(1):20. doi: 10.1186/s13072-025-00584-8.
In Drosophila, architectural proteins are frequently found in promoters, including those of genes with extremely high expression levels, such as ribosomal protein genes (RPGs). The involvement of several of these proteins in gene regulation in Drosophila has been shown, but the exact mechanisms of their possible cooperative action have not been fully elucidated.
In this study we dissected the contribution of the architectural proteins Opbp and M1BP, which are co-localized at several RPG promoters near the transcription start site, to promoter functioning. We found that Opbp has two domains that directly interact with CP190, Putzig (Pzg), and Chromator (Chro) proteins, the cofactors which are required for the activation of housekeeping (hk) gene promoters. These domains have redundant functions in vivo and can tether the cofactors forming open chromatin regions when are artificially recruited to the "closed" chromatin. Additionally, we observed interactions between M1BP and the same cofactors. In the transgene assay, the transcription driven by the 192-bp part of Rpl27A RPG promoter is fully dependent on the presence of at least one Opbp or M1BP binding site and it is sufficient for the very high activity of this promoter integrated into the hk gene cluster and moderate expression outside the cluster, while presence of both sites even more facilitates transcription.
This study demonstrates that different architectural proteins can work independently and in cooperation and fulfill partially redundant functions in the activation of RPG promoters.
在果蝇中,结构蛋白经常出现在启动子中,包括那些具有极高表达水平的基因的启动子,如核糖体蛋白基因(RPGs)。已经证明其中几种蛋白质参与果蝇的基因调控,但其可能的协同作用的确切机制尚未完全阐明。
在本研究中,我们剖析了在转录起始位点附近的几个RPG启动子上共定位的结构蛋白Opbp和M1BP对启动子功能的贡献。我们发现Opbp有两个结构域,它们直接与CP190、Putzig(Pzg)和Chromator(Chro)蛋白相互作用,这些辅助因子是管家(hk)基因启动子激活所必需的。这些结构域在体内具有冗余功能,当人工招募到“封闭”染色质时,它们可以束缚形成开放染色质区域的辅助因子。此外,我们观察到M1BP与相同的辅助因子之间存在相互作用。在转基因试验中,由Rpl27A RPG启动子的192bp部分驱动的转录完全依赖于至少一个Opbp或M1BP结合位点的存在,并且对于整合到hk基因簇中的该启动子的极高活性以及在基因簇外的适度表达而言是足够的,而两个位点的存在甚至更有利于转录。
本研究表明,不同的结构蛋白可以独立发挥作用并协同工作,在RPG启动子的激活中发挥部分冗余功能。