Emilia Modolo Pinto1, Enilze M.S.F. Ribeiro2, Jinling Wang1, Aaron H. Phillips3, Richard W. Kriwacki3 and Gerard P. Zambetti1 1Department of Pathology, St. Jude Children's Research Hospital, Memphis, Tennessee 38105, USA; 2Programa de Pós-graduação em Genética, Universidade Federal do Paraná, Curitiba, Paraná, 81531-980, Brazil; 3Department of Structural Biology, St. Jude Children's Research Hospital, Memphis, Tennessee 38105, USA Corresponding authors: emilia.pintostjude.org; enilzeribeirogmail.com; gerard.zambettistjude.org Abstract

TP53 plays a critical role as a tumor suppressor by controlling cell cycle progression, DNA repair, and apoptosis. Post-translational modifications such as acetylation of specific lysine residues in the DNA binding and carboxy-terminus regulatory domains modulate its tumor suppressor activities. In this study, we addressed the functional consequences of the germline TP53 p.K164E (NM_000546.5: c.490A>G) variant identified in a patient with early-onset breast cancer and a significant family history of cancer. K164 is a conserved residue located in the L2 loop of the p53 DNA binding domain that is post-translationally modified by acetylation. In silico, in vitro, and in vivo analyses demonstrated that the glutamate substitution at K164 marginally destabilizes the p53 protein structure but significantly impairs sequence-specific DNA binding, transactivation, and tumor cell growth inhibition. Although p.K164E is currently considered a variant of unknown significance by different clinical genetic testing laboratories, the clinical and laboratory-based findings presented here provide strong evidence to reclassify TP53 p.K164E as a likely pathogenic variant.

Received September 12, 2023. Accepted November 27, 2023.