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Insights into a nonhomologous integration pathway in the dermatophyte Trichophyton mentagrophytes : efficient targeted gene disruption by use of mutants lacking ligase IV
Author(s) -
Alshahni Mohamed Mahdi,
Yamada Tsuyoshi,
Takatori Kosuke,
Sawada Takuo,
Makimura Koichi
Publication year - 2011
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/j.1348-0421.2010.00283.x
Subject(s) - biology , trichophyton , mutant , dna ligase , homologous recombination , non homologous end joining , genetics , dna , dna repair , gene , dermatophyte , locus (genetics) , microbiology and biotechnology , antifungal
Targeted gene disruption experiments in Trichophyton mentagrophytes are impeded by the dominant of repair of DNA double strand breaks through a nonhomologous end joining pathway (NHEJ). Inactivation of human DNA ligase IV homologs, which is involved in the final step of the NHEJ pathway, has been shown to enhance homologous recombination (HR) frequency in filamentous fungi. To improve the frequency of HR in T. mentagrophytes , the lig4 homolog ( TmLIG4 ) was disrupted. T. mentagrophytes lacking TmLIG4 showed no discernable phenotypic differences when compared to wild‐type controls. Both mutant and parent strains had almost identical growth ability, sporulation rate and sensitivity to DNA damaging agents. When four different loci were disrupted in the TMLIG4‐deficient mutant, HR frequencies reached as high as 93% depending on the locus, whereas they ranged from 0%–40% in the wild‐type. These results suggest that studies in strains lacking TmLIG4 would help to improve our understanding of dermatophytosis by facilitating the genetic manipulation of dermatophytes.