Ramón Ailyn C, Basukala Om, Massimi Paola, Thomas Miranda, Perera Yasser, Banks Lawrence, Perea Silvio E
Molecular Oncology Group, Department of Pharmaceuticals, Biomedical Research Division, Center for Genetic Engineering and Biotechnology (CIGB), Havana 10600, Cuba.
Tumor Virology Group, International Centre for Genetic Engineering and Biotechnology (ICGEB), AREA Science Park, 34149 Trieste, Italy.
Viruses. 2022 Jul 30;14(8):1681. doi: 10.3390/v14081681.
CIGB-300 is a clinical-grade anti-Protein Kinase CK2 peptide, binding both its substrate's phospho-acceptor site and the CK2α catalytic subunit. The cyclic p15 inhibitory domain of CIGB-300 was initially selected in a phage display library screen for its ability to bind the CK2 phospho-acceptor domain ofHPV-16 E7. However, the actual role of this targeting in CIGB-300 antitumoral mechanism remains unexplored. Here, we investigated the physical interaction of CIGB-300 with HPV-E7 and its impact on CK2-mediated phosphorylation. Hence, we studied the relevance of targeting E7 phosphorylation for the cytotoxic effect induced by CIGB-300. Finally, co-immunoprecipitation experiments followed by western blotting were performed to study the impact of the peptide on the E7-pRB interaction. Interestingly, we found a clear binding of CIGB-300 to the N terminal region of E7 proteins of the HPV-16 type. Accordingly, the in vivo physical interaction of the peptide with HPV-16 E7 reduced CK2-mediated phosphorylation of E7, as well as its binding to the tumor suppressor pRB. However, the targeting of E7 phosphorylation by CIGB-300 seemed to be dispensable for the induction of cell death in HPV-18 cervical cancer-derived C4-1 cells. These findings unveil novel molecular clues to the means by which CIGB-300 triggers cell death in cervical cancer cells.
CIGB - 300是一种临床级抗蛋白激酶CK2肽,它既能结合其底物的磷酸化受体位点,又能结合CK2α催化亚基。CIGB - 300的环状p15抑制结构域最初是在噬菌体展示文库筛选中因其能够结合HPV - 16 E7的CK2磷酸化受体结构域而被选中的。然而,这种靶向作用在CIGB - 300抗肿瘤机制中的实际作用仍未得到探索。在此,我们研究了CIGB - 300与HPV - E7的物理相互作用及其对CK2介导的磷酸化的影响。因此,我们研究了靶向E7磷酸化与CIGB - 300诱导的细胞毒性作用的相关性。最后,进行了免疫共沉淀实验,随后进行蛋白质印迹分析,以研究该肽对E7 - pRB相互作用的影响。有趣的是,我们发现CIGB - 300与16型HPV的E7蛋白的N末端区域有明显的结合。相应地,该肽与HPV - 16 E7在体内的物理相互作用降低了CK2介导的E7磷酸化及其与肿瘤抑制因子pRB的结合。然而,CIGB - 300对E7磷酸化的靶向作用似乎对于在HPV - 18宫颈癌来源的C4 - 1细胞中诱导细胞死亡是可有可无的。这些发现揭示了CIGB - 300在宫颈癌细胞中触发细胞死亡的新分子线索。