PMID- 3653252
OWN - NLM
STAT- MEDLINE
DCOM- 19871105
LR  - 20190707
IS  - 0014-4827 (Print)
IS  - 0014-4827 (Linking)
VI  - 172
IP  - 1
DP  - 1987 Sep
TI  - In situ factors affecting stability of the DNA helix in interphase nuclei and 
      metaphase chromosomes.
PG  - 168-79
AB  - The data from earlier cytochemical studies, in which the metachromatic 
      fluorochrome acridine orange (AO) was used to differentially stain single vs 
      double-stranded DNA, suggested that DNA in situ in intact metaphase chromosomes 
      or in condensed chromatin of G0 cells is more sensitive to denaturation, induced 
      by heat or acid, than DNA in decondensed chromatin of interphase nuclei. Present 
      studies show that, indeed, DNA in permeabilized metaphase cells, in contrast to 
      cells in interphase, when exposed to buffers of low pH (1.5-2.8) becomes 
      digestible with the single-strand-specific S1 or mung bean nucleases. A variety 
      of extraction procedures and enzymatic treatments provided evidence that the 
      presence of histones, HMG proteins, and S-S bonds in chromatin, as well as 
      phosphorylation or poly(ADP)ribosylation of chromatin proteins, can be excluded 
      as a factor responsible for the differential sensitivity of metaphase vs 
      interphase DNA to denaturation. Cell treatment with NaCl at a concentration of 
      1.2 N and above abolished the difference between interphase and mitotic cells, 
      rendering DNA in mitotic cells less sensitive to denaturation; such treatment 
      also resulted in decondensation of chromatin visible by microscopy. The present 
      data indicate that structural proteins extractable with greater than or equal to 
      1.2 N NaCl may be involved in anchoring DNA to the nuclear matrix or chromosome 
      scaffold and may be responsible for maintaining a high degree of chromatin 
      compaction in situ, such as that observed in metaphase chromosomes or in G0 
      cells. Following dissociation of histones, the high spatial density of the 
      charged DNA polymer may induce topological strain on the double helix, thus 
      decreasing its local stability; this can be detected by metachromatic staining of 
      DNA with AO or digestion with single-strand-specific nucleases.
FAU - Darzynkiewicz, Z
AU  - Darzynkiewicz Z
AD  - Sloan-Kettering Institute for Cancer Research, Walker Laboratory, Rye, New York 
      10580.
FAU - Traganos, F
AU  - Traganos F
FAU - Carter, S P
AU  - Carter SP
FAU - Higgins, P J
AU  - Higgins PJ
LA  - eng
GR  - CA-23296/CA/NCI NIH HHS/United States
GR  - CA-28704/CA/NCI 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 (DNA, Neoplasm)
RN  - EN464416SI (Ethidium)
RN  - F30N4O6XVV (Acridine Orange)
SB  - IM
MH  - Acridine Orange
MH  - Animals
MH  - Cell Cycle
MH  - Cell Nucleus/*ultrastructure
MH  - Chromosomes/*ultrastructure
MH  - DNA, Neoplasm/*metabolism
MH  - Ethidium
MH  - Interphase
MH  - Leukemia L1210/*pathology
MH  - Metaphase
MH  - Mice
MH  - Nucleic Acid Conformation
EDAT- 1987/09/01 00:00
MHDA- 1987/09/01 00:01
CRDT- 1987/09/01 00:00
PHST- 1987/09/01 00:00 [pubmed]
PHST- 1987/09/01 00:01 [medline]
PHST- 1987/09/01 00:00 [entrez]
AID - 0014-4827(87)90103-0 [pii]
AID - 10.1016/0014-4827(87)90103-0 [doi]
PST - ppublish
SO  - Exp Cell Res. 1987 Sep;172(1):168-79. doi: 10.1016/0014-4827(87)90103-0.