Matrix attachment region (MAR) properties and abnormal expansion of AT island minisatellites in FRA16B fragile sites in leukemic CEM cells.
AT-rich minisatellites (AT islands) are sites of genomic instability in cancer cells and targets for extremely lethal AT-specific drugs, such as bizelesin. Here we investigated the AT islands in the FRA16B fragile site region for their possible roles in the organization of DNA on the nuclear matrix. The FRA16B AT island nominally spans approximately 3 kb of mostly >90% A/T DNA. In silico analysis indicates that this domain exhibits characteristics of nuclear matrix attachment regions (MARs): an exceptionally intense computed 'MAR potential' and profound duplex destabilization and flexibility. FRA16B repeats specifically bind to isolated nuclear matrices, which indicates their in vitro MAR function. This binding is several-fold greater than that of a known MAR in the c-myc gene. AT islands in fragile sites FRA16B and FRA16D are significantly more abundant in CEM cells that are hypersensitive to bizelesin compared to normal WI-38 cells. FRA16B overabundance in CEM is due to an approximately 10-fold expansion of FRA16B repeats. The expanded FRA16B minisatellites in CEM cells preferentially localize to the nuclear matrix-associated DNA indicating their in vivo MAR function. The unexpanded repeats in WI-38 cells localize to the loop DNA. The c-myc MAR is also matrix-associated in CEM cells while localizing to loop DNA in WI-38 cells. These results are the first to demonstrate that AT islands in fragile sites can function as MARs both in vitro and in vivo. The ability of FRA16B-mediated MAR sites to rearrange depending on the repeat expansion status could be relevant to both genomic instability of cancer cells and their sensitivity to AT-island targeting drugs.
Department of Radiation Oncology, University of Texas Health Science Center, 14960 Omicron Drive, San Antonio, TX 78245, USA.
Jennifer A
JA
Jackson
Alex V
AV
Trevino
Maryanne C
MC
Herzig
Terence S
TS
Herman
Jan M
JM
Woynarowski
D020856:AT Rich Sequence / Q000235:genetics*; D001665:Binding Sites; D045744:Cell Line, Tumor; D002873:Chromosome Fragility / Q000235:genetics*; D002885:Chromosomes, Human, Pair 16 / Q000235:genetics; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D043243:DNA Sequence, Unstable / Q000235:genetics*; D000080890:Duocarmycins; D016259:Genes, myc / Q000235:genetics; D005914:Globins / Q000235:genetics; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D007938:Leukemia / Q000235:genetics*; D018598:Minisatellite Repeats / Q000235:genetics*; D008969:Molecular Sequence Data; D015530:Nuclear Matrix / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction; D014508:Urea / Q000031:analogs & derivatives* / Q000494:pharmacology
D000080890:Duocarmycins; D007211:Indoles; D014508:Urea; D005914:Globins; D004247:DNA; C072257:bizelesin