A methylated human 9-kb repetitive sequence on acrocentric chromosomes is homologous to a subtelomeric repeat in chimpanzees.
We have implemented an approach for the detection of DNA alterations in cancer by means of computerized analysis of end-labeled genomic fragments, separated in two dimensions. Analysis of two-dimensional patterns of neuroblastoma tumors, prepared by first digesting DNA with the methylation-sensitive restriction enzyme Not I, yielded a multicopy fragment which was detected in some tumor patterns but not in normal controls. Cloning and sequencing of the fragment, isolated from two-dimensional gels, yielded a sequence with a strong homology to a subtelomeric sequence in chimpanzees and which was previously reported to be undetectable in humans. Fluorescence in situ hybridization indicated the occurrence of this sequence in normal tissue, for the most part in the satellite regions of acrocentric chromosomes. A product containing this sequence was obtained by telomere-anchored PCR using as a primer an oligonucleotide sequence from the cloned fragment. Our data suggest demethylation of cytosines at the cloned Not I site and in neighboring DNA in some tumors, compared with normal tissue, and suggest a greater similarity between human and chimpanzee subtelomeric sequences than was previously reported.
Thoraval
D
D
Department of Pediatrics, University of Michigan School of Medicine, Ann Arbor 48109, USA.
Asakawa
J
J
Kodaira
M
M
Chang
C
C
Radany
E
E
Kuick
R
R
Lamb
B
B
Richardson
B
B
Neel
J V
JV
Glover
T
T
Hanash
S
S
D000818:Animals; D001483:Base Sequence; D005075:Biological Evolution; D015139:Blotting, Southern; D002874:Chromosome Mapping; D002877:Chromosomes, Human; D004247:DNA / Q000096:biosynthesis / Q000737:chemistry; D017931:DNA Primers; D004273:DNA, Neoplasm / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D015186:Hominidae / Q000235:genetics*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008214:Lymphocytes / Q000166:cytology; D008745:Methylation; D008969:Molecular Sequence Data; D009447:Neuroblastoma / Q000235:genetics / Q000378:metabolism; D002679:Pan troglodytes / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012091:Repetitive Sequences, Nucleic Acid
D017931:DNA Primers; D004273:DNA, Neoplasm; D004247:DNA