DNA repair mechanisms affecting cytotoxicity by streptozotocin in E. coli.
Mechanisms underlying cytotoxicity by the monofunctional nitrosourea streptozotocin (STZ) were evaluated in DNA repair-deficient E. coli mutants. Strains not proficient in recombinational repair which lack either RecA protein or RecBC gene products were highly sensitive to STZ. In contrast, cells that constitutively synthesize RecA protein and cannot initiate SOS repair mechanisms because of uncleavable LexA repressor (recAo98 lexA3) were resistant to this drug compared to a lexA3 strain. Further, E. coli cells lacking both 3-methyladenine DNA glycosylases I (tag) and II (alkA) also were highly sensitive to STZ. DNA synthesis was most inhibited by STZ in recA and alkA tag E. coli mutants, but was suppressed less markedly in wild-type and recBC cells. DNA degradation was most extensive in recA E. coli after STZ treatment, while comparable in recBC, alkA tag, and wild-type cells. Although increased single-stranded DNA breaks were present after STZ treatment in recA and recBC mutants compared to the wild type, no significant increase in DNA single-stranded breaks was noted in alkA tag E. coli. Further, DNA breaks in recBC cells were repaired, while those present in recA cells were not. These findings establish the critical importance of both recombinational repair and 3-methyladenine DNA glycosylase in ameliorating cytotoxic effects and DNA damage caused by STZ in E. coli.
Fram
R J
RJ
Department of Medicine, University of Massachusetts Medical School, Worcester 01655.
Mack
S L
SL
George
M
M
Marinus
M G
MG
D001426:Bacterial Proteins / Q000096:biosynthesis; D004260:DNA Repair; D004261:DNA Replication / Q000187:drug effects; D004269:DNA, Bacterial / Q000096:biosynthesis / Q000187:drug effects; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D005838:Genotype; D012329:RNA, Bacterial / Q000096:biosynthesis; D013045:Species Specificity; D013311:Streptozocin / Q000494:pharmacology*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D012329:RNA, Bacterial; D013311:Streptozocin