Unraveling novel mRNA transcripts of the human DNA N-glycosylase 1 (NTHL1) gene with the implementation of an innovative targeted DNA-seq assay.

Autor: Diamantopoulos MA; Department of Biochemistry and Molecular Biology, National and Kapodistrian University of Athens, Athens, Greece., Adamopoulos PG; Department of Biochemistry and Molecular Biology, National and Kapodistrian University of Athens, Athens, Greece., Tsiakanikas P; Department of Biochemistry and Molecular Biology, National and Kapodistrian University of Athens, Athens, Greece., Nisotakis T; Department of Biochemistry and Molecular Biology, National and Kapodistrian University of Athens, Athens, Greece., Skourou PC; Department of Biochemistry and Molecular Biology, National and Kapodistrian University of Athens, Athens, Greece., Scorilas A; Department of Biochemistry and Molecular Biology, National and Kapodistrian University of Athens, Athens, Greece. Electronic address: ascorilas@biol.uoa.gr.
Jazyk: angličtina
Zdroj: Gene [Gene] 2024 Dec 20; Vol. 930, pp. 148856. Date of Electronic Publication: 2024 Aug 13.
DOI: 10.1016/j.gene.2024.148856
Abstrakt: The human NTHL1 gene encodes a DNA glycosylase that plays a key role in the base excision repair (BER) pathway, repairing oxidative DNA damage and maintaining genome integrity. The physiological activity of NTHL1 is crucial in preventing genetic alterations that can lead to cancer. In this study, we employed an innovative targeted DNA sequencing (DNA-seq) methodology to explore the transcriptional landscape of the NTHL1 gene, revealing previously uncharacterized alternative splicing events and novel exons. Our designed approach provided significantly improved sequencing depth and coverage, enabling the identification of novel NTHL1 mRNA transcripts. Bioinformatics analysis confirmed all annotated splice junctions of the main NTHL1 transcripts (v.1 - v.3) and revealed novel mRNA transcripts (NTHL1 v.4 - v.9) derived from splicing events between annotated exons as well as mRNAs containing previously uncharacterized exons (NTHL1 v.10 - v.14). Quantitative PCR analysis highlighted a diverse expression pattern of these novel transcripts across different human cell lines, suggesting cell-specific roles and regulatory mechanisms. Notably, NTHL1 v.5 was overexpressed in luminal A breast cancer cells (MCF-7), while v.13 was prominent in triple negative (BT-20), HER2 + breast cancer (SK-BR-3), prostate, colorectal cancer cells and HEK-293 cells. Our findings suggest that specific novel NTHL1 transcripts may encode protein isoforms with distinct structural features, as indicated by ribosome profiling datasets, while others containing premature termination codons could function as long non-coding RNAs. These insights enhance our understanding of NTHL1 regulatory role and its potential as a biomarker and therapeutic target in human malignancies. This study underscores the importance of exploring the transcriptional diversity of NTHL1 to fully elucidate its role in cancer pathobiology.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2024. Published by Elsevier B.V.)
Databáze: MEDLINE
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