Insights into a nonhomologous integration pathway in the dermatophyte Trichophyton mentagrophytes: efficient targeted gene disruption by use of mutants lacking ligase IV
Article first published online: 22 DEC 2010
© 2010 The Societies and Blackwell Publishing Asia Pty Ltd
Microbiology and Immunology
Volume 55, Issue 1, pages 34–43, January 2011
How to Cite
Alshahni, M. M., Yamada, T., Takatori, K., Sawada, T. and Makimura, K. (2011), Insights into a nonhomologous integration pathway in the dermatophyte Trichophyton mentagrophytes: efficient targeted gene disruption by use of mutants lacking ligase IV. Microbiology and Immunology, 55: 34–43. doi: 10.1111/j.1348-0421.2010.00283.x
- Issue published online: 22 DEC 2010
- Article first published online: 22 DEC 2010
- Accepted manuscript online: 29 OCT 2010 12:05PM EST
- Received 24 June 2010; revised 9 September 2010; 28 September 2010; accepted 21 October 2010.
- gene targeting;
- ligase IV;
- nonhomologous integration
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.