Photocrosslinking in ruthenium-labelled duplex oligonucleotides.
The formation of a photoadduct between a [Ru(1,4,5,8-tetraazaphenanthrene)(2)4,7-diphenylphenanthroline](2+) complex chemically attached to a synthetic oligonucleotide, and a guanine moiety in a complementary targeted single-stranded DNA molecule was studied for ten 17-mer duplexes by denaturing gel electrophoresis. This photoadduct formation leads to photocrosslinking of the two strands. The percentage quenching of luminescence of the complex by electron transfer was compared to the resulting yield of photocrosslinked product. This yield does not only depend on the ionisation potential of the guanine bases, which are electron donors, but also on other factors, such as the position of the guanine bases as compared to the site of attachment of the complex. The photocrosslinking yield is higher when the guanine moieties are towards the 3' end on the complementary strand as compared to the tethering site. Computer modelling results are in agreement with this preference for the 3' side for the photoreaction. Interestingly, the photocrosslink is not alkali labile. Moreover, a type III exonuclease enzyme is blocked at the position of photocrosslinking.
Lentzen
O
O
Université Libre de Bruxelles, Organic Chemistry and Photochemistry, CP. 160/08, 50 Avenue F. D. Roosevelt, Belgium.
Constant
J-F
JF
Defrancq
E
E
Prévost
M
M
Schumm
S
S
Moucheron
C
C
Dumy
P
P
Kirsch-De Mesmaeker
A
A
D003432:Cross-Linking Reagents / Q000528:radiation effects*; D004579:Electron Transport; D006147:Guanine; D007553:Isotope Labeling; D009841:Oligonucleotides / Q000737:chemistry*; D010777:Photochemistry; D012429:Ruthenium Radioisotopes
D003432:Cross-Linking Reagents; D009841:Oligonucleotides; D012429:Ruthenium Radioisotopes; D006147:Guanine