PMID- 2461311
OWN - NLM
STAT- MEDLINE
DCOM- 19881230
LR  - 20190707
IS  - 0014-4827 (Print)
IS  - 0014-4827 (Linking)
VI  - 179
IP  - 2
DP  - 1988 Dec
TI  - Staining with pyronin Y detects changes in conformation of RNA during mitosis and 
      hyperthermia of CHO cells.
PG  - 535-44
AB  - Cellular RNA in Chinese hamster ovary (CHO) cells synchronized in mitosis (M) or 
      G2 phase, as well as in interphase cells subjected to hyperthermia (42 degrees C, 
      10 min), was stained with acridine orange (AO), ethidium bromide (EB), or pyronin 
      Y (PY) and the resultant fluorescence was measured by flow cytometry. Total RNA 
      content detected after staining with AO increased in M as compared to G2-phase 
      cells, consistent with continued RNA synthesis during G2 phase. The content of 
      double-stranded RNA, stained with EB (after DNase treatment), was also somewhat 
      higher in M cells. In contrast, the stainability of RNA with PY decreased by 27% 
      in M- compared to G2-phase cells. Furthermore, a decrease in stainability of RNA 
      with PY was observed in G2 cells compared to cells in G1 phase. In separate 
      experiments, RNA stainability with AO or EB was generally unaffected when 
      interphase CHO cells were exposed to 42 degrees C for 10 min, though this same 
      treatment resulted in a 26% decrease in RNA stainability with PY. The decreased 
      PY stainability of cellular RNA in M or heat-treated cells was observed at a 
      relatively narrow range of dye concentration (1.0-2.0 micrograms/ml). The 
      observed hypochromicity of RNA coincides with dissociation of polyribosomes into 
      single ribosomes known to occur during mitosis and following exposure to 
      hyperthermia. It is presumed that the phenomenon involves selective denaturation 
      and condensation of ribosomal (r) RNA by PY in single ribosomes which does not 
      occur in polyribosomes. While the molecular mechanisms responsible for 
      stabilization of rRNA in polyribosomes preventing its denaturation and 
      condensation by PY are unknown, PY appears to be a sensitive probe that can be 
      used to detect and study these changes in rRNA confirmation in situ.
FAU - Traganos, F
AU  - Traganos F
AD  - Sloan Kettering Institute for Cancer Research, New York, New York 10021.
FAU - Crissman, H A
AU  - Crissman HA
FAU - Darzynkiewicz, Z
AU  - Darzynkiewicz Z
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Exp Cell Res
JT  - Experimental cell research
JID - 0373226
RN  - 0 (Xanthenes)
RN  - 63231-63-0 (RNA)
RN  - W659G165T1 (Pyronine)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Cricetinae
MH  - *Hot Temperature
MH  - Mitosis
MH  - *Nucleic Acid Conformation
MH  - Pyronine/*metabolism
MH  - RNA/*analysis
MH  - Staining and Labeling
MH  - Xanthenes/*metabolism
EDAT- 1988/12/01 00:00
MHDA- 1988/12/01 00:01
CRDT- 1988/12/01 00:00
PHST- 1988/12/01 00:00 [pubmed]
PHST- 1988/12/01 00:01 [medline]
PHST- 1988/12/01 00:00 [entrez]
AID - 10.1016/0014-4827(88)90291-1 [doi]
PST - ppublish
SO  - Exp Cell Res. 1988 Dec;179(2):535-44. doi: 10.1016/0014-4827(88)90291-1.