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Mutation Research, 42 (1977) 33-44
© Elsevier/North-Holland Biomedical Press
MUTAGENIC DNA REPAIR IN ESCHERICHIA COLI. V. MUTATION
FREQUENCY DECLINE AND ERROR·FREE POST-REPLICATION
REPAIR IN AN EXCISION-PROFICIENT STRAIN
M.H.L. GREEN, B.A. BRIDGES, J.E. EYFJORD and W.J. MURIEL
M.R.C. Cell Mutation Unit, University of Sussex, Folmer, Brighton BN 1 9QG (Breat Britain)
(Received April 12th, 1976)
(Revision received July 15th, 1976)
(Accepted September 9th, 1976)
Summary
Mutation frequency decline (MFD) is an irreversible loss of newly-induced
suppressor mutations occurring in excision-proficient Escherichia coli during a
short period of incubation in minimal medium before plating on broth- or Casa-
mino acids-enriched selective agar. It is known that MFD of UV-induced muta-
tions may occur before DNA containing pre-mutagenic lesions is replicated, but
we conclude that MFD can also occur after the damaged DNA has been repli-
cated on the basis of the following evidence. (1) Mutation fixation in rich me-
dium (i.e., loss of susceptibility to mutation frequency decline) with ethyl
methanesulphonate mutagenesis begins immediately, whereas with UV it is de-
layed for 20-30 min. (2) The delay in mutation fixation after UV can be ex-
plained neither by inhibition of DNA replication nor by a delay in the appear-
ance of error-prone repair activity in the irradiated population. (3) MFD at
later times after UV irradiation is more rapid and is less strongly inhibited by
caffeine than is MFD immediately after irradiation. (4) Excision is virtually
complete 20 min after 3 J m? UV but at that time virtually all mutations are
still susceptible to MFD.
We have presented evidence elsewhere that in bacteria there is an alternative
error-free excision-dependent type of post-replication repair of potentially
mutagenic daughter strand gaps. We suggest that this process is inhibited at
tRN A loci in the presence of nutrient broth or Casamino acids, possibly be-
cause of a broth-dependent change in the structure of the single-stranded region
including the tRNA locus.
Introduction
When Escherichia coli WP2 is exposed to ultraviolet light (UV) and assayed
for the induction of Trp+ mutants the yield is 5 to 10 times greater if the plates