Study of IFI16 protein interaction with G4 prone DNA sequences and its influence on p53 transcription activity

abstract

English

The IFI16 protein interactions with DNA are important in many biological processes such as regulation of replication, transcription and translation. The binding properties of IFI16 to G4 prone sequences and the effect of this bond on reporter gene transcription, induced by the well-known tumor suppressor p53, were studied in a yeast isogenic system. The G4 prone DNA sequences, with the p53 PUMA binding site (derived from the PUMA gene promoter), were located upstream of a minimal gene promoter LUC1. The KSHV sequence derived from the G4 herpesvirus KSHV and its adenine-substituted mutants KSHV-1NO and KSHV-3NO were analyzed by CD spectroscopy. The binding of the IFI16 protein to these sequences was studied by EMSA. The CD spectroscopy results showed the formation of parallel G4 of KSHV and KSHV-1NO sequences in potassium ion-containing buffer. In vitro competitive EMSA demonstrated IFI16 protein interactions with CD retrieved G4 formation sequences, without significant binding to PUMA or KSHV-3NO oligonucleotides. In vivo experiments showed a repression of LUC1 transcription due to stabilization of KSHV by IFI16 protein and an increase of LUC1 transcription due to p53 guidance to its binding sites by IFI16 protein in the case of absence of any IFI16 binding site.

IFI16, p53, G-quarduplex, DNA-protein interactions, protein-protein interactions, transcription activity, Yeasts one-hybrid isogenic systém, EMSA, CD spectroscopy

KRATOCHVILOVÁ, L.; VALKOVÁ, N.; BRÁZDA, V.

20. 12. 2023

2564-615X

The EuroBiotech Journal

7 (2023)

s1

Republic of Italy

38

39

2

https://sciendo.com/article/10.2478/ebtj-2023-0019