RNA-DNA hybrids containing damaged DNA are substrates for RNase H.
During the replication of the lagging strand, RNA-DNA hybrids are formed and the RNA is subsequently degraded by the action of RNase H. Little is known about the effects of damaged DNA on lagging strand replication and subsequent RNA removal. The rates and sites of digestion by E. coli RNase H of RNA-DNA hybrids containing either a thymine glycol or urea site in the DNA strand have been examined. The cleavage patterns for duplexes containing thymine glycol or urea differ from that of a fully complementary duplex. There is one major product of the digestion of the fully complementary hybrid, but three products are formed in the reactions with the hybrids containing damaged DNAs. Cleavage is partially redirected to the position adjacent to the damaged sites. The overall rate of cleavage of these hybrids containing damaged DNA is comparable to that of the fully complementary duplex. These results indicate that the cleavage of RNA-DNA hybrids by RNase H is less selective when a damaged site is present in the DNA strand.
Shiels
J C
JC
Chemistry Department, Wesleyan University, Middletown, CT 06459, USA.
Jerkovic
B
B
Baranger
A M
AM
Bolton
P H
PH
D004247:DNA / Q000378:metabolism*; D004249:DNA Damage; D004926:Escherichia coli / Q000201:enzymology*; D009692:Nucleic Acid Heteroduplexes / Q000378:metabolism*; D012313:RNA / Q000378:metabolism*; D016914:Ribonuclease H / Q000378:metabolism*
D009692:Nucleic Acid Heteroduplexes; D012313:RNA; D004247:DNA; D016914:Ribonuclease H