Anticancer peptide CIGB-300 binds to nucleophosmin/B23, impairs its CK2-mediated phosphorylation, and leads to apoptosis through its nucleolar disassembly activity.
CIGB-300, formerly known as P15-tat, is a proapoptotic peptide with established antiproliferative activity in vitro and antitumoral activity in vivo. This hypothesis-driven peptide was initially selected for its ability to impair the in vitro CK2-mediated phosphorylation in one of its substrates through direct binding to the conserved acidic phosphoaceptor domain. However, the actual in vivo target(s) on human cancer cells among the hundreds of CK2 substrates as well as the subsequent events that lead to apoptosis on tumor cells remains to be determined. In this work, we identified the multifunctional oncoprotein nucleophosmin/B23 as a major target for CIGB-300. In vivo, the CIGB-300-B23 interaction was shown by pull-down experiments and confirmed by the early in situ colocalization of both molecules in the cell nucleolus. Moreover, CIGB-300 inhibits the CK2-mediated phosphorylation of B23 in a dose-dependent fashion both in vitro and in vivo as shown using the recombinant GST fusion protein and the metabolic labeling approach, respectively. Such phosphorylation impairment was correlated with the ability of CIGB-300 to induce nucleolar disassembly as documented by the use of established markers for nucleolar structure. Finally, we showed that such a sequence of events leads to the rapid and massive onset of apoptosis both at the molecular and cellular levels. Collectively, these findings provide important clues by which the CIGB-300 peptide exerts its proapoptotic effect on tumor cells and highlights the suitability of the B23/CK2 pathway for cancer-targeted therapy.
Perera
Yasser
Y
Laboratory of Molecular Oncology, Division of Pharmaceuticals, Center for Genetic Engineering and Biotechnology, PO Box 6162, Havana CP10600, Cuba. yasser.perera@cigb.edu.cu
Farina
Hernán G
HG
Gil
Jeovanis
J
Rodriguez
Arielis
A
Benavent
Fernando
F
Castellanos
Lila
L
Gómez
Roberto E
RE
Acevedo
Boris E
BE
Alonso
Daniel F
DF
Perea
Silvio E
SE
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D014408:Biomarkers, Tumor; D047390:Casein Kinase II / Q000378:metabolism*; D045744:Cell Line, Tumor; D002466:Cell Nucleolus / Q000378:metabolism*; D006801:Humans; D042541:Intracellular Space; D058990:Molecular Targeted Therapy; D009687:Nuclear Proteins / Q000378:metabolism*; D000090243:Nucleophosmin; D010456:Peptides, Cyclic / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology*; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects*; D011485:Protein Binding; D021381:Protein Transport; D055752:Small Cell Lung Carcinoma / Q000473:pathology
D000970:Antineoplastic Agents; D014408:Biomarkers, Tumor; C000716967:NPM1 protein, human; D009687:Nuclear Proteins; D010456:Peptides, Cyclic; D010750:Phosphoproteins; C036825:phosphoprotein C23; D000090243:Nucleophosmin; D047390:Casein Kinase II; C532510:CIGB-300