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Photocrosslinking of G-Quadruplex-Forming Sequences found in Human Promoters
Jillian E Smith-Carpenter1, John-Stephen Taylor1
1Department of Chemistry, Washington University, St. Louis, MO.
Photochemistry and Photobiology
|August 8, 2018
Summary
Human promoter DNA G-quadruplexes can form UV-induced crosslinks, similar to telomeric DNA. This study identified specific promoter sequences forming nonadjacent cyclobutane pyrimidine dimers (CPDs), suggesting G-quadruplexes as potential in vivo probes.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Human telomeric DNA G-quadruplexes are known to form UV-induced crosslinks.
- Over 40% of human promoter sequences contain putative G-quadruplex-forming regions.
- UV light can induce cyclobutane pyrimidine dimer (CPD) formation in G-quadruplex structures.
Purpose of the Study:
- To investigate the potential of promoter G-quadruplexes to form UV-induced crosslinks.
- To identify specific promoter sequences capable of forming G-quadruplexes and undergoing UV photocrosslinking.
- To characterize the types of CPDs formed in promoter G-quadruplexes.
Main Methods:
- In vitro UVB irradiation of 15 putative G-quadruplex-forming promoter sequences.
- Mass spectrometry to confirm CPD formation.
- Circular dichroism, melting temperature analysis, and chemical footprinting to analyze G-quadruplex structures.
Main Results:
- Three promoter sequences were confirmed to form nonadjacent CPDs upon UVB irradiation.
- Both nonadjacent T=T CPDs and a novel nonadjacent T=U CPD (from deamination of T=C CPD) were identified.
- Structural analyses confirmed the presence of G-quadruplexes in the studied sequences.
Conclusions:
- Promoter G-quadruplexes can form UV-induced nonadjacent CPDs in vitro.
- These findings suggest promoter G-quadruplexes could serve as in vivo probes for non-B DNA structures.
- In vivo formation of these crosslinks may have significant biological implications.
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