Sep 29, 2019

Infrared nanospectroscopic mapping of a single metaphase chromosome

Nucleic Acids Research
Ewelina LipiecWojciech M Kwiatek

Abstract

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]2py2] 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, the...Continue Reading

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Mentioned in this Paper

Antineoplastic Agents
Genome
Interphase
DNA Methylation
HOOC-CH2-Cha-Pro-Arg-(CH2-O-CH2-CF3)
Metaphase
Cell Nucleus
Gene Expression
Microscopy, Atomic Force
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