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KRATOCHVILOVÁ, L. VALKOVÁ, N. BRÁZDA, V.
Original Title
Study of IFI16 protein interaction with G4 prone DNA sequences and its influence on p53 transcription activity
Type
abstract
Language
English
Original Abstract
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.
Keywords
IFI16, p53, G-quarduplex, DNA-protein interactions, protein-protein interactions, transcription activity, Yeasts one-hybrid isogenic systém, EMSA, CD spectroscopy
Authors
KRATOCHVILOVÁ, L.; VALKOVÁ, N.; BRÁZDA, V.
Released
20. 12. 2023
ISBN
2564-615X
Periodical
The EuroBiotech Journal
Year of study
7 (2023)
Number
s1
State
Republic of Italy
Pages from
38
Pages to
39
Pages count
2
URL
https://sciendo.com/article/10.2478/ebtj-2023-0019
BibTex
@misc{BUT185685, author="KRATOCHVILOVÁ, L. and VALKOVÁ, N. and BRÁZDA, V.", title="Study of IFI16 protein interaction with G4 prone DNA sequences and its influence on p53 transcription activity", year="2023", journal="The EuroBiotech Journal", volume="7 (2023)", number="s1", pages="38--39", doi="10.2478/ebtj-2023-0019", issn="2564-615X", url="https://sciendo.com/article/10.2478/ebtj-2023-0019", note="abstract" }