EVIDENCE FOR INTRAMOLECULARLY FOLDED I-DNA STRUCTURES IN BIOLOGICALLY RELEVANT CCC-REPEAT SEQUENCES

被引:191
作者
MANZINI, G [1 ]
YATHINDRA, N [1 ]
XODO, LE [1 ]
机构
[1] UNIV MADRAS,DEPT CRYSTALLOG & BIOPHYS,MADRAS 600025,TAMIL NADU,INDIA
关键词
D O I
10.1093/nar/22.22.4634
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The structural behaviour of repetitive cytosine DNA is examined in the oligodeoxynucleotide sequences of (CCCTAA)(3)CCCT (HTC4), GC(TCCC)(3)TCCT(TCCC)3 (KRC6) and the methylated (CCCT)(3)TCCT(CCCT)(3)C (KRM6) by circular dichroism (CD), gel electrophoresis (PAGE), and ultra violet (UV) absorbance studies. Ail the three sequences exhibit a pH-induced cooperative structural transition as monitored by CD. An intense positive CD band around 285 nm develops on lowering the pH from 8 to slightly acidic condition, indicative of the formation of base pairs between protonated cytosines. The oligomers are found to melt in a fully reversible and cooperative fashion, with a melting temperature (T-m) of around 50 degrees C at pH 5.5. The melting temperatures are independent from DNA concentration, indicative of an intramolecular process involved in the structural formation. PAGE experiments performed with P-32-labeled samples as well as with normal staining procedures show a predominantly single band migration for all the three oligomers suggestive of a unimolecular structure. From pH titrations the number of protons required for generating the structures formed by HTC4, KRC6 and KRM6 results to be around six. These findings strongly suggest that all the three sequences adopt an intramolecular i-motif structure. The demonstration of i-motif structure for KRC6, a critical functional stretch of the c-ki-ras promoter proto-oncogene, besides the human telomeric sequence HTC4, may be suggestive of larger significance in the functioning of DNA.
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页码:4634 / 4640
页数:7
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