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Published on: March 18, 2010
Effects of mutagen-sensitive mus mutations on spontaneous mitotic recombination in Aspergillus
Abstract:
Methyl methane-sulfonate (MMS)-sensitive, radiation-induced mutants of Aspergillus were shown to define nine new DNA repair genes, musK to musS. To test mus mutations for effects on mitotic recombination, intergenic crossing over was assayed between color markers and their centromeres, and intragenic recombination between two distinguishable adE alleles. Of eight mutants analyzed, four showed significant deviations from mus+ controls in both tests. Two mutations, musK and musL, reduced recombination, while musN and musQ caused increases. In contrast, musO diploids produced significantly higher levels only for intragenic recombination. Effects were relatively small, but averages between hypo- and hyperrec mus differed 15-20-fold. In musL diploids, most of the rare color segregants resulted from mitotic malsegregation rather than intergenic crossing over. This indicates that the musL gene product is required for recombination and that DNA lesions lead to chromosome loss when it is deficient. In addition, analysis of the genotypes of intragenic (ad+) recombinants showed that the musL mutation specifically reduced single allele conversion but increased complex conversion types (especially recombinants homozygous for ad+). Similar analysis revealed differences between the effects of two hyperrec mutations; musN apparently caused high levels solely of mitotic crossing over, while musQ increased various conversion types but not reciprocal crossovers. These results suggest that mitotic gene conversion and crossing over, while generally associated, are affected differentially in some of the mus strains of Aspergillus nidulans.
Insights
New DNA repair genes (musK-musS) in Aspergillus affect DNA repair and recombination. Some mutations decrease recombination, while others increase it, impacting gene conversion and crossing over differently.
Area of Science:
- Genetics
- Molecular Biology
- DNA Repair
Background:
- Radiation-induced mutants in Aspergillus nidulans are crucial for understanding DNA repair pathways.
- Methyl methanesulfonate (MMS)-sensitive mutants define genes involved in DNA damage response.
Purpose of the Study:
- To investigate the role of newly identified DNA repair genes (musK-musS) in mitotic recombination.
- To determine how mutations in these genes affect intergenic crossing over and intragenic recombination.
Main Methods:
- Assaying intergenic crossing over between color markers and centromeres in Aspergillus mutants.
- Analyzing intragenic recombination between distinguishable adE alleles.
- Genotyping intragenic recombinants to differentiate conversion types.
Main Results:
- Four of eight mus mutations significantly altered recombination frequencies.
- musK and musL mutations reduced recombination, while musN and musQ increased it.
- musL deficiency led to chromosome loss instead of recombination, and differential effects on allele conversion were observed. musN and musQ showed distinct impacts on crossing over and gene conversion.
Conclusions:
- The musL gene product is essential for recombination, and its deficiency results in chromosome loss upon DNA damage.
- Mitotic gene conversion and crossing over are differentially regulated and can be independently affected by specific DNA repair gene mutations.
- These findings highlight the complex interplay between DNA repair and recombination pathways in Aspergillus.
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