title: Three DUF1996 Proteins Localize in Vacuoles and Function in Fungal Responses to Multiple Stresses and Metal Ions.
abstract: Many annotated fungal genomes harbour high proportions of hypothetical proteins with or without domains of unknown function (DUF). Here, three novel proteins (342-497 amino acids), each containing only a single large DUF1996 (231-250 residues) region with highly conserved head (DPIXXP) and tail (HXDXXXGW) signatures, were expressed as eGFP-tagged fusion proteins and shown to specifically localize in the vacuoles of Beauveria bassiana, a filamentous fungal entomopathogen; therefore, these proteins were named vacuole-localized proteins (VLPs). The VLPs have one to three homologues in other entomopathogenic or non-entomopathogenic filamentous fungi but no homologues in yeasts. The large DUF1996 regions can be formulated as D-X4-P-X5-6-H-X-H-X3-G-X25-26-D-X-S-X-YW-X-P-X123-203-CP-X39-48-H-X-D-X3-GW; the identical residues likely involve in a proton antiport system for intracellular homeostasis. Single deletions of three VLP-coding genes (vlp1-3) increased fungal sensitivities to cell wall perturbation, high osmolarity, oxidation, and several metal ions. Conidial thermotolerance decreased by ~11% in two Δvlp mutants, and UV-B resistance decreased by 41-57% in three Δvlp mutants. All the changes were restored by targeted gene complementation. However, the deletions did not influence fungal growth, conidiation, virulence or Cu(2+) sensitivity. Our findings unveiled a role for the DUF1996 regions of three B. bassiana VLPs in the regulation of multiple stress responses and environmental adaptation.
authors.affiliation: Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058, People's Republic of China.
authors.forename: Sen-Miao
authors.initials: SM
authors.lastname: Tong
authors.affiliation: Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058, People's Republic of China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058, People's Republic of China.
authors.forename: Sheng-Hua
authors.initials: SH
authors.lastname: Ying
authors.affiliation: Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058, People's Republic of China.
authors.forename: Ming-Guang
authors.initials: MG
authors.lastname: Feng
mesh_terms: D052982:Beauveria / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005816:Genetic Complementation Test; D010802:Phylogeny; D000072417:Protein Domains; D013312:Stress, Physiological; D013696:Temperature; D014617:Vacuoles / Q000378:metabolism*; D037521:Virulence Factors
chemical_list: D005656:Fungal Proteins; D037521:Virulence Factors
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