Journal
NUCLEIC ACIDS RESEARCH
Volume 47, Issue 18, Pages -Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkz630
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Funding
- National Science Center of the Republic of Poland (Narodowe Centrum Nauki--NCN) [2014/13/D/NZ1/01014]
- CRUS Sciex Project [13.295]
- Swiss National Science Foundation (SNF) [P2ELP2 162116, P300P2 171219]
- Malopolska Regional Operational Programme Measure 5.1 Krakow Metropolitan Area [MRPO.05.01.00-12-013/15]
- PL-Grid Infrastructure
- Marian Smoluchowski Krakow Scientific Consortium 'Mater-Energy-Future' [KNOW]
- Darwin College, United Kingdom
- AustralianResearchCouncil (ARC) Future Fellowship [FT120100926]
- Institute of Nuclear Physics, Polish Academy of Sciences
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The integrity of the chromatin structure is essential to every process occurring within eukaryotic nuclei. However, there are no reliable tools to decipher the molecular composition of metaphase chromosomes. Here, we have applied infrared nanospectroscopy (AFM-IR) to demonstrate molecular difference between eu- and heterochromatin and generate infrared maps of single metaphase chromosomes revealing detailed information on their molecular composition, with nanometric lateral spatial resolution. AFM-IR coupled with principal component analysis has confirmed that chromosome areas containing euchromatin and heterochromatin are distinguishable based on differences in the degree of methylation. AFM-IR distribution of eu- and heterochromatin was compared to standard fluorescent staining. We demonstrate the ability of our methodology to locate spatially the presence of anticancer drug sites in metaphase chromosomes and cellular nuclei. We show that the anticancer 'rule breaker' platinum compound [Pt[N(p-HC6F4)CH2](2)py(2)] preferentially binds to heterochromatin, forming localized discrete foci due to condensation of DNA interacting with the drug. Given the importance of DNA methylation in the development of nearly all types of cancer, there is potential for infrared nanospectroscopy to be used to detect gene expression/suppression sites in the whole genome and to become an early screening tool for malignancy.
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