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Title: Stem-loop structures of the repetitive DNA sequences located at human centromeres

Conference ·
OSTI ID:10102555
; ; ;  [1]; ; ;  [2];  [1]
  1. Los Alamos National Lab., NM (United States)
  2. California Univ., Davis, CA (United States). Dept. of Biological Chemistry

The presence of the highly conserved repetitive DNA sequences in the human centromeres argues for a special role of these sequences in their biological functions - most likely achieved by the formation of unusual structures. This prompted us to carry out quantitative one- and two-dimensional nuclear magnetic resonance (lD/2D NMR) spectroscopy to determine the structural properties of the human centromeric repeats, d(AATGG){sub n.d}(CCATT){sub n}. The studies on centromeric DNAs reveal that the complementary sequence, d(AATGG){sub n.d}(CCATT){sub n}, adopts the usual Watson-Crick B-DNA duplex and the pyrimidine-rich d(CCATT){sub n} strand is essentially a random coil. However, the purine-rich d(AATGG){sub n} strand is shown to adopt unusual stem-loop structures for repeat lengths, n=2,3,4, and 6. In addition to normal Watson-Crick A{center_dot}T pairs, the stem-loop structures are stabilized by mismatch A{center_dot}G and G{center_dot}G pairs in the stem and G-G-A stacking in the loop. Stem-loop structures of d(AATGG)n are independently verified by gel electrophoresis and nuclease digestion studies. Thermal melting studies show that the DNA repeats, d(AATGG){sub n}, are as stable as the corresponding Watson-Crick duplex d(AATGG){sub n.d}(CCATT){sub n}. Therefore, the sequence d(AATGG){sub n} can, indeed, nucleate a stem-loop structure at little free-energy cost and if, during mitosis, they are located on the chromosome surface they can provide specific recognition sites for kinetochore function.

Research Organization:
Los Alamos National Lab., NM (United States)
Sponsoring Organization:
USDOE, Washington, DC (United States)
DOE Contract Number:
W-7405-ENG-36
OSTI ID:
10102555
Report Number(s):
LA-UR-93-3073; CONF-9306237-2; ON: DE93040187
Resource Relation:
Conference: 8. conversation in the discipline biomolecular stereodynamics,Albany, NY (United States),Jun 1993; Other Information: PBD: [1993]
Country of Publication:
United States
Language:
English

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