Brh2-Dss1 interplay enables properly controlled recombination in Ustilago maydis.
Brh2, the BRCA2 homolog in Ustilago maydis, functions in recombinational repair of DNA damage by regulating Rad51 and is, in turn, regulated by Dss1. Dss1 is not required for Brh2 stability in vivo, nor for Brh2 to associate with Rad51, but is required for formation of green fluorescent protein (GFP)-Rad51 foci following DNA damage by gamma radiation. To understand more about the interplay between Brh2 and Dss1, we isolated mutant variants of Brh2 able to bypass the requirement for Dss1. These variants were found to lack the entire C-terminal DNA-Dss1 binding domain but to maintain the N-terminal region harboring the Rad51-interacting BRC element. GFP-Rad51 focus formation was nearly normal in brh2 mutant cells expressing a representative Brh2 variant with the C-terminal domain deleted. These findings suggest that the N-terminal region of Brh2 has an innate ability to organize Rad51. Survival after DNA damage was almost fully restored by a chimeric form of Brh2 having a DNA-binding domain from RPA70 fused to the Brh2 N-terminal domain, but Rad51 focus formation and mitotic recombination were elevated above wild-type levels. The results provide evidence for a mechanism in which Dss1 activates a Brh2-Rad51 complex and balances a finely regulated recombinational repair system.
Kojic
Milorad
M
Department of Microbiology and Immunology, Box 62, Cornell University Weill Medical College,1300 York Ave., New York, NY 10021, USA.
Zhou
Qingwen
Q
Lisby
Michael
M
Holloman
William K
WK
D002467:Cell Nucleus / Q000378:metabolism; D049109:Cell Proliferation; D015825:Chromosomes, Fungal; D004249:DNA Damage; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D009154:Mutation / Q000235:genetics; D011485:Protein Binding; D051135:Rad51 Recombinase; D011995:Recombination, Genetic / Q000235:genetics*; D014588:Ustilago / Q000166:cytology / Q000235:genetics* / Q000378:metabolism* / Q000528:radiation effects
D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D051135:Rad51 Recombinase