Triplex-forming DNAs in the human interphase nucleus visualized in situ by polypurine/polypyrimidine DNA probes and antitriplex antibodies

Ohno Mizuki, Tatsuo Fukagawa, Jeremy S. Lee, Toshimichi Ikemura

Research output: Contribution to journalArticle

60 Citations (Scopus)

Abstract

The polypurine/polypyrimidine (PuPy) tracts present in the human genome are known to be scattered among and within chromosomes. In PuPy tract sequences, triplex formation occurs readily under physiological conditions, leaving single-stranded DNAs capable of hybridization with complementary single-stranded DNAs and RNAs. The formation of single-strands and transmolecular triplexes is thought to enable sequences spaced distantly along the genome to associate with each other and organize nuclear DNA into ordered configurations. Triplex-forming DNAs in the human interphase nucleus were analyzed by combining fluorescence in situ "non-denaturing" hybridization employing PuPy tract probes and immunodetection by antitriplex antibodies. The non-denaturing hybridization technique, which has been used to detect RNA, may detect single-stranded DNAs in non-denatured nuclei, if present. Probes such as (GA/TC)n and (GAA/TTC)n sequences gave sequence-specific signals that overlapped with or were closely associated with triplexes immunolocalized by using known antitriplex antibodies. Pretreatment of nuclei with antitriplex antibodies blocked probe signal formation. Signal formation was resistant to pretreatment of nuclei with RNases but sensitive to single strand-specific nucleases. Triplexes visualized differentially with distinct PuPy tract probes were associated spatially with centromeric sequences in the interphase nucleus in a sequence-specific manner.

Original languageEnglish
Pages (from-to)201-213
Number of pages13
JournalChromosoma
Volume111
Issue number3
DOIs
Publication statusPublished - Jan 1 2002

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Single-Stranded DNA
Interphase
DNA Probes
Antibodies
RNA
Human Genome
Ribonucleases
Protein Sorting Signals
Complementary DNA
Chromosomes
Fluorescence
Genome
DNA
triplex DNA

All Science Journal Classification (ASJC) codes

  • Genetics
  • Genetics(clinical)

Cite this

Triplex-forming DNAs in the human interphase nucleus visualized in situ by polypurine/polypyrimidine DNA probes and antitriplex antibodies. / Mizuki, Ohno; Fukagawa, Tatsuo; Lee, Jeremy S.; Ikemura, Toshimichi.

In: Chromosoma, Vol. 111, No. 3, 01.01.2002, p. 201-213.

Research output: Contribution to journalArticle

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