title: Monitoring the reversible B to A-like transition of DNA in eukaryotic cells using Fourier transform infrared spectroscopy.
abstract: The ability to detect DNA conformation in eukaryotic cells is of paramount importance in understanding how some cells retain functionality in response to environmental stress. It is anticipated that the B to A transition might play a role in resistance to DNA damage such as heat, desiccation and toxic damage. To this end, conformational detail about the molecular structure of DNA has been derived primarily from in vitro experiments on extracted or synthetic DNA. Here, we report that a B- to A-like DNA conformational change can occur in the nuclei of intact cells in response to dehydration. This transition is reversible upon rehydration in air-dried cells. By systematically monitoring the dehydration and rehydration of single and double-stranded DNA, RNA, extracted nuclei and three types of eukaryotic cells including chicken erythrocytes, mammalian lymphocytes and cancerous rodent fibroblasts using Fourier transform infrared (FTIR) spectroscopy, we unequivocally assign the important DNA conformation marker bands within these cells. We also demonstrate that by applying FTIR spectroscopy to hydrated samples, the DNA bands become sharper and more intense. This is anticipated to provide a methodology enabling differentiation of cancerous from non-cancerous cells based on the increased DNA content inherent to dysplastic and neoplastic tissue.
authors: Center for Biospectroscopy and School of Chemistry, Monash Immunology and Stem Cell Laboratories, Monash University, Clayton, Victoria, 3800 Australian Synchrotron, 800 Blackburn Road, Clayton, Victoria 3168, Australia.
authors: Donna R
authors: DR
authors: Whelan
authors: Keith R
authors: KR
authors: Bambery
authors: Philip
authors: P
authors: Heraud
authors: Mark J
authors: MJ
authors: Tobin
authors: Max
authors: M
authors: Diem
authors: Don
authors: D
authors: McNaughton
authors: Bayden R
authors: BR
authors: Wood
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D004247:DNA / Q000737:chemistry*; D044704:DNA, A-Form / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000737:chemistry; D004912:Erythrocytes / Q000737:chemistry; D005347:Fibroblasts / Q000737:chemistry; D008214:Lymphocytes / Q000737:chemistry; D051379:Mice; D009690:Nucleic Acid Conformation; D025341:Principal Component Analysis; D012313:RNA / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D044704:DNA, A-Form; D004277:DNA, Single-Stranded; D012313:RNA; D004247:DNA