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Genetic effects of N-methyl-N′-nitro-N-nitrosoguanidine in Neurospora crassa

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Summary

The mutagenicity of N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) was studied with a genetically marked, balanced heterokaryon of Neurospora crassa. Types of genetic alterations detectable in this system are (a) point mutations in the ad-3 A and ad-3 B loci, (b) multilocus (chromosome) deletions in the ad-3 region, and (c) recessive lethal mutations in the whole genome. Study of the inactivation kinetics of the heterokaryotic and homokaryotic conidial fractions makes it possible to distinguish between nuclear and cytoplasmic inactivation.

Forward mutations in the ad-3 region induced by MNNG in the heterokaryotic fraction of conidia were obtained by a direct method, with the following results: (1) forward-mutation frequency increases as the square of the time of treatment, (2) MNNG is an extremely efficient mutagen, e. g., the frequency of mutation in the ad-3 region (2 loci) was 0.14% after 240 min treatment with 25 μM MNNG at pH 7.0 with 73.4% survival, (3) at least 98.1% of the MNNG-induced ad-3 mutants are point mutations, (4) tests for genotype and allelic complementation showed that (a) the frequency of genotypes was ad-3 A=19.7%, ad-3 B=80.3% and ad-3 A ad-3 B=0.0%, and (b) 81.8% of the ad-3 B mutants have allelic complementation with 79.9% nonpolarized and 1.9% polarized complementation patterns and 18.2% noncomplementing mutants, and (5) the ration between mutations in the ad-3 A and ad-3 B loci and spectrum of complementation patterns among the ad-3 B mutants was independent of dose. Comparison of the spectrum of the complementation patterns among ad-3 B mutants induced by MNNG with the spectrum among ad-3 B mutants induced by 2-aminopurine, nitrous acid, hydroxylamine, and the acridine mustard derivative ICR-170 suggests that the majority of the MNNG-induced mutants have guanine-cytosine at the mutant site.

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Abbreviations

GC:

guanine-cytosine

MNNG:

N-methyl-N′-nitro-N-nitrosoguanidine

HA:

hydroxylamine

AT:

adenine-thymine

MMS:

methyl methanesulfonate

2AP:

2-aminopurine

ICR-170:

acridine mustard derivative-(2-methoxy-6-chloro-9-[(ethyl-2-chloroethyl) amino propylamino] acridine dihydrochloride)

NA:

nitrous acid

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Authors and Affiliations

  1. Biology Division, Oak Ridge National Laboratory, Oak Ridge, Tennessee

    H. V. Malling & F. J. De Serres

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  1. H. V. Malling
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  2. F. J. De Serres
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Communicated by F. Kaudewitz

Research sponsored by the U.S. Atomic Energy Commission under contract with the Union Carbide Corporation.

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Malling, H.V., De Serres, F.J. Genetic effects of N-methyl-N′-nitro-N-nitrosoguanidine in Neurospora crassa . Molec. Gen. Genetics 106, 195–207 (1970). https://doi.org/10.1007/BF00340379

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