Abstract
Fapy·dG is produced in DNA as a result of oxidative stress. Under some conditions Fapy·dG is formed in greater yields than 8-oxodG from a common chemical precursor. Recently, Fapy·dG and its C-nucleoside analogue were incorporated in chemically synthesized oligonucleotides at defined sites. Like 8-oxodG, Fapy·dG instructs DNA polymerase to misincorporate dA opposite it in vitro. The interactions of DNA containing Fapy·dG or the nonhydrolyzable analogue with Fpg and MutY are described. Fpg excises Fapy·dG (KM = 2.0 nM, kcat = 0.14 min-1) opposite dC ∼17-fold more efficiently than when mispaired with dA, which is misinserted by DNA polymerase in vitro. Fpg also prefers to bind duplexes containing Fapy·dG·dC or β-C-Fapy·dG·dC compared to those in which the lesion is opposite dA. MutY incises dA when it is opposite Fapy·dG and strongly binds duplexes containing the lesion or β-C-Fapy· dG. Incision from Fapy·dG·dA is faster than from dG·dA mispairs but slower than from DNA containing 8-oxodG opposite dA. These data demonstrate that Fapy·dG closely resembles the interactions of 8-oxodG with two members of the GO repair pathway in vitro. The similar effects of Fapy·dG and 8-oxodG on DNA polymerase and repair enzymes in vitro raise the question as to whether Fapy·dG elicits similar effects in vivo.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 9755-9760 |
| Number of pages | 6 |
| Journal | Biochemistry |
| Volume | 42 |
| Issue number | 32 |
| DOIs | |
| State | Published - Aug 19 2003 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Biochemistry
Cite this
Repair of DNA containing Fapy·dG and its β-C-nucleoside analogue by formamidopyrimidine DNA glycosylase and MutY. / Wiederholt, Carissa J.; Delaney, Michael O.; Pope, Mary Ann; David, Sheila S.; Greenberg, Marc M.
In: Biochemistry, Vol. 42, No. 32, 19.08.2003, p. 9755-9760.Research output: Contribution to journal › Article
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TY - JOUR
T1 - Repair of DNA containing Fapy·dG and its β-C-nucleoside analogue by formamidopyrimidine DNA glycosylase and MutY
AU - Wiederholt, Carissa J.
AU - Delaney, Michael O.
AU - Pope, Mary Ann
AU - David, Sheila S.
AU - Greenberg, Marc M.
PY - 2003/8/19
Y1 - 2003/8/19
N2 - Fapy·dG is produced in DNA as a result of oxidative stress. Under some conditions Fapy·dG is formed in greater yields than 8-oxodG from a common chemical precursor. Recently, Fapy·dG and its C-nucleoside analogue were incorporated in chemically synthesized oligonucleotides at defined sites. Like 8-oxodG, Fapy·dG instructs DNA polymerase to misincorporate dA opposite it in vitro. The interactions of DNA containing Fapy·dG or the nonhydrolyzable analogue with Fpg and MutY are described. Fpg excises Fapy·dG (KM = 2.0 nM, kcat = 0.14 min-1) opposite dC ∼17-fold more efficiently than when mispaired with dA, which is misinserted by DNA polymerase in vitro. Fpg also prefers to bind duplexes containing Fapy·dG·dC or β-C-Fapy·dG·dC compared to those in which the lesion is opposite dA. MutY incises dA when it is opposite Fapy·dG and strongly binds duplexes containing the lesion or β-C-Fapy· dG. Incision from Fapy·dG·dA is faster than from dG·dA mispairs but slower than from DNA containing 8-oxodG opposite dA. These data demonstrate that Fapy·dG closely resembles the interactions of 8-oxodG with two members of the GO repair pathway in vitro. The similar effects of Fapy·dG and 8-oxodG on DNA polymerase and repair enzymes in vitro raise the question as to whether Fapy·dG elicits similar effects in vivo.
AB - Fapy·dG is produced in DNA as a result of oxidative stress. Under some conditions Fapy·dG is formed in greater yields than 8-oxodG from a common chemical precursor. Recently, Fapy·dG and its C-nucleoside analogue were incorporated in chemically synthesized oligonucleotides at defined sites. Like 8-oxodG, Fapy·dG instructs DNA polymerase to misincorporate dA opposite it in vitro. The interactions of DNA containing Fapy·dG or the nonhydrolyzable analogue with Fpg and MutY are described. Fpg excises Fapy·dG (KM = 2.0 nM, kcat = 0.14 min-1) opposite dC ∼17-fold more efficiently than when mispaired with dA, which is misinserted by DNA polymerase in vitro. Fpg also prefers to bind duplexes containing Fapy·dG·dC or β-C-Fapy·dG·dC compared to those in which the lesion is opposite dA. MutY incises dA when it is opposite Fapy·dG and strongly binds duplexes containing the lesion or β-C-Fapy· dG. Incision from Fapy·dG·dA is faster than from dG·dA mispairs but slower than from DNA containing 8-oxodG opposite dA. These data demonstrate that Fapy·dG closely resembles the interactions of 8-oxodG with two members of the GO repair pathway in vitro. The similar effects of Fapy·dG and 8-oxodG on DNA polymerase and repair enzymes in vitro raise the question as to whether Fapy·dG elicits similar effects in vivo.
UR - http://www.scopus.com/inward/record.url?scp=0042527365&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0042527365&partnerID=8YFLogxK
U2 - 10.1021/bi034844h
DO - 10.1021/bi034844h
M3 - Article
C2 - 12911318
AN - SCOPUS:0042527365
VL - 42
SP - 9755
EP - 9760
JO - Biochemistry
JF - Biochemistry
SN - 0006-2960
IS - 32
ER -