DNA stability

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(Deamination)
 
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* Single stranded DNA deaminates significantly faster (> 100 times) than double stranded DNA
* Single stranded DNA deaminates significantly faster (> 100 times) than double stranded DNA
* Mispaired Cs (C:C and T:C) are 1-2 orders of magnitude more likely to deaminate than in double stranded DNA (similar to single stranded DNA)
* Mispaired Cs (C:C and T:C) are 1-2 orders of magnitude more likely to deaminate than in double stranded DNA (similar to single stranded DNA)
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* The rate of deamination of C mispairs was around <math>(8-40)\cdot 10^{-10}</math>/sec at 60C and at 37C, it is about <math>(4-13)\cdot 10^{-10}</math>/sec
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* The rate of deamination of C mispairs is around <math>(8-40)\cdot 10^{-10}</math>/sec at 60C and about <math>(4-13)\cdot 10^{-10}</math>/sec at 37C
==Strand cleavage==
==Strand cleavage==
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#lindahl72b pmid=4559796
#lindahl72b pmid=4559796
</biblio>
</biblio>
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[[Category:DNA]]

Current revision

Although DNA is generally viewed as a stable molecule, many conditions can cause loss of DNA bases or strand breakage.

Contents

Depurination

  • Depurination involves the loss of purine bases forming abasic sites
  • Depurination is one of the two limiting factors in chemical synthesis of long DNA oligos (the other is coupling efficiency)
  • DNA under physiological conditions has been estimated to depurinate at a rate of 3\cdot 10^{-11}/sec at 37C and pH 7.4 [1]
  • Heating DNA for 10m@100 at pH 7.0 leads to about 1 apurinic site per 1000 base pairs
  • The activation energy of depurination is around 29 kcal/mol
  • Higher temperatures lead to faster depurination
  • Denatured DNA depurinates at about 4 times the rate of dsDNA @ pH 7.4
  • Methylated As (6-methyladenine) found in bacteria are depurinated 4 times faster than the unmethylated purine bases
  • Depurination decreases at higher pH (thus acidic conditions favor depurination)
  • Depurination proceeds more rapidly in buffers of low ionic strength
  • Depurination is correlated with lower transformation efficiency
  • Depurination is independent of sequence

Deamination

  • Cytosine can be spontaneously deaminated to form uracil.
  • Cytosine in native DNA is estimated to deanimate with a rate constant of 7\cdot 10^{-13}/sec at 37C and 10 − 10/sec at 70C
  • Single stranded DNA deaminates significantly faster (> 100 times) than double stranded DNA
  • Mispaired Cs (C:C and T:C) are 1-2 orders of magnitude more likely to deaminate than in double stranded DNA (similar to single stranded DNA)
  • The rate of deamination of C mispairs is around (8-40)\cdot 10^{-10}/sec at 60C and about (4-13)\cdot 10^{-10}/sec at 37C

Strand cleavage

  • Under physiological conditions (pH 7.4, ionic strength of 150mM with 10mM Mg++ ions), the lifetime of a phosphodiester bond at an abasic site is 190 hours @ 37C [2]
  • Abasic sites are alkali-labile. Under mildly alkaline conditions, β-elimination occurs which nicks 3' to the abasic site leaving a 5'-P on the downstream fragment
  • Under strong alkaline conditions, δ-elimination will occur after β-elimination which completely removes the abasic site leaving a 3'-P on the upstream fragment and a 5'-P on the downstream fragment
  • Amines (such as amino acids or polyamines like putrescine, spermidiine, and spermine) greatly increase the rate of strand breakage at abasic sites

References

  1. Lindahl T and Nyberg B. . pmid:4626532. PubMed HubMed [lindahl72]
  2. Lindahl T and Andersson A. . pmid:4559796. PubMed HubMed [lindahl72b]
  3. Frederico LA, Kunkel TA, and Shaw BR. . pmid:8329382. PubMed HubMed [frederico93]
All Medline abstracts: PubMed HubMed
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