PMID- 6195003
OWN - NLM
STAT- MEDLINE
DCOM- 19831217
LR  - 20190707
IS  - 0014-4827 (Print)
IS  - 0014-4827 (Linking)
VI  - 148
IP  - 1
DP  - 1983 Oct
TI  - Denaturation of RNA and DNA in situ induced by acridine orange.
PG  - 31-46
AB  - The products of interaction of acridine orange (AO) with single-stranded (ss) 
      nucleic acids are precipitates which exhibit red luminescence. Titration of rRNA 
      or thymus DNA with AO results in formation of such products suggesting that the 
      dye, per se, denatures double-stranded (ds) sections of these biopolymers. This 
      transition, measured as the increase of red luminescence, a concomitant decrease 
      of green fluorescence, and followed by an increase of light scatter of the 
      AO-nucleic acid complexes, is cooperative and at 0.15 N NaCl occurs at 4-20 and 
      10-50 microM range of AO concentration for rRNA and DNA, respectively. The 
      changes in stainability of nucleic acids in situ, in permealized cells, occur at 
      higher AO concentration. Thus, the transition of RNA in situ is biphasic and seen 
      at 20-120 microM AO. In the presence of EDTA, however, the change is monophasic 
      and shifted to the 10-30 microM range of AO concentration. The change in 
      stainability of DNA also shows two phases: one at 30-60 microM and another at 
      70-120 microM of AO. Extraction of basic proteins with 0.08 N HCl shifts the 
      transition of DNA to the 30-60 microM AO concentration and makes it monophasic. 
      The observed differences in denaturability of RNA vs DNA explain the specificity 
      of AO in differential staining of these bipolymers in histochemical reactions. In 
      living cells the products of interaction of AO with nucleic acids are detected by 
      electron microscopy. In the cytoplasm of interphase cells the formation of dense 
      precipitates within ribosomes and polysomes, simultaneous with a specific 
      retraction of ribosome-polysome complexes from the periphery of the cell to the 
      nucleus is evident. The latter suggests higher order organization of these 
      particles involving their association with each other or with the nucleus via 
      polyanionic macromolecules which collapse upon binding with AO. The DNA in 
      heterochromatin is more sensitive to AO-induced denaturation, as evidenced by the 
      fact that the dense complexes are formed preferentially in the regions of 
      condensed chromatin of the interphase nucleus, or in metaphase chromosomes.
FAU - Darzynkiewicz, Z
AU  - Darzynkiewicz Z
FAU - Evenson, D
AU  - Evenson D
FAU - Kapuscinski, J
AU  - Kapuscinski J
FAU - Melamed, M R
AU  - Melamed MR
LA  - eng
GR  - CA28704/CA/NCI NIH HHS/United States
GR  - ES3035/ES/NIEHS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Exp Cell Res
JT  - Experimental cell research
JID - 0373226
RN  - 0 (Heterochromatin)
RN  - 0 (RNA, Ribosomal)
RN  - 63231-63-0 (RNA)
RN  - 9007-49-2 (DNA)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange/*pharmacology
MH  - Animals
MH  - Cell Line
MH  - *DNA
MH  - Heterochromatin/analysis
MH  - Interphase
MH  - Leukemia, Experimental
MH  - Metaphase
MH  - Mice
MH  - Microscopy, Electron
MH  - Nucleic Acid Denaturation/*drug effects
MH  - *RNA
MH  - RNA, Ribosomal
MH  - Ribosomes/analysis
MH  - Staining and Labeling
EDAT- 1983/10/01 00:00
MHDA- 1983/10/01 00:01
CRDT- 1983/10/01 00:00
PHST- 1983/10/01 00:00 [pubmed]
PHST- 1983/10/01 00:01 [medline]
PHST- 1983/10/01 00:00 [entrez]
AID - 0014-4827(83)90185-4 [pii]
AID - 10.1016/0014-4827(83)90185-4 [doi]
PST - ppublish
SO  - Exp Cell Res. 1983 Oct;148(1):31-46. doi: 10.1016/0014-4827(83)90185-4.