Cytosine methylation is a widespread epigenetic regulation mechanism. In healthy mature cells, methylation occurs at CpG dinucleotides within promoters, where it primarily silences gene expression by modifying the binding affinity of transcription factors to the promoters. Conversely, a recent study showed that in stem cells and cancer cell precursors, methylation also occurs at non-CpG pairs and involves introns and even gene bodies. The epigenetic role of such methylations and the molecular mechanisms by which they induce gene regulation remain elusive. The topology of both physiological and aberrant non-CpG methylation patterns still has to be detailed and could be revealed by using the differential stability of the duplexes formed between site-specific oligonucleotide probes and the corresponding methylated regions of genomic DNA. Here, we present a systematic study of the thermal stability of a DNA oligonucleotide sequence as a function of the number and position of non-CpG methylation sites. The melting temperatures were determined by monitoring the fluorescence of donor-acceptor duallabelled oligonucleotides at various temperatures. An empirical model that estimates the methylationinduced variations in the standard values of hybridization entropy and enthalpy was developed.

Nardo, L., Lamperti, M., Salerno, D., Cassina, V., Missana, N., Bondani, M., et al. (2015). Effects of non-CpG site methylation on DNA thermal stability: A fluorescence study. NUCLEIC ACIDS RESEARCH, 43(22), 10722-10733 [10.1093/nar/gkv884].

Effects of non-CpG site methylation on DNA thermal stability: A fluorescence study

NARDO, LUCA
;
SALERNO, DOMENICO;CASSINA, VALERIA;MISSANA, NATALIA;TEMPESTINI, ALESSIA
Penultimo
;
MANTEGAZZA, FRANCESCO
Ultimo
2015

Abstract

Cytosine methylation is a widespread epigenetic regulation mechanism. In healthy mature cells, methylation occurs at CpG dinucleotides within promoters, where it primarily silences gene expression by modifying the binding affinity of transcription factors to the promoters. Conversely, a recent study showed that in stem cells and cancer cell precursors, methylation also occurs at non-CpG pairs and involves introns and even gene bodies. The epigenetic role of such methylations and the molecular mechanisms by which they induce gene regulation remain elusive. The topology of both physiological and aberrant non-CpG methylation patterns still has to be detailed and could be revealed by using the differential stability of the duplexes formed between site-specific oligonucleotide probes and the corresponding methylated regions of genomic DNA. Here, we present a systematic study of the thermal stability of a DNA oligonucleotide sequence as a function of the number and position of non-CpG methylation sites. The melting temperatures were determined by monitoring the fluorescence of donor-acceptor duallabelled oligonucleotides at various temperatures. An empirical model that estimates the methylationinduced variations in the standard values of hybridization entropy and enthalpy was developed.
Articolo in rivista - Articolo scientifico
DNA; Fluorometry; Nucleic Acid Denaturation; Oligonucleotide Probes; Temperature; Thermodynamics; DNA Methylation; Genetics
English
2015
43
22
10722
10733
none
Nardo, L., Lamperti, M., Salerno, D., Cassina, V., Missana, N., Bondani, M., et al. (2015). Effects of non-CpG site methylation on DNA thermal stability: A fluorescence study. NUCLEIC ACIDS RESEARCH, 43(22), 10722-10733 [10.1093/nar/gkv884].
File in questo prodotto:
Non ci sono file associati a questo prodotto.

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/135584
Citazioni
  • Scopus 24
  • ???jsp.display-item.citation.isi??? 21
Social impact