Reference - Detail
|Nakamura T, Abe H, Hirata A, Shimoda C.
|ADAM family protein Mde10 is essential for development of spore envelopes in the fission yeast Schizosaccharomyces pombe.
We report the identification of Schizosaccharomyces pombe mde10+ as a gene possessing a FLEX element, which forms a binding site for the meiosis-specific transcription factor Mei4. In fact, mde10+ is transcribed only in diploid cells that are induced to meiosis in a Mei4-dependent manner. Western blot analysis indicated that the epitope-tagged Mde10 protein accumulates transiently during meiosis and then rapidly decreases. Mde10 is a multidomain protein containing a metalloprotease catalytic domain, a disintegrin domain, a cysteine-rich domain, and membrane-spanning regions, all of which are shared by members of the mammalian ADAM family. A fusion protein of Mde10 and green fluorescent protein localized to the endoplasmic reticulum during meiosis and was located at the peripheral region of spores at the end of meiosis. An mde10Delta deletion mutant showed no apparent defects in meiosis, sporulation, or spore germination. However, the mutant spores exhibited an aberrant surface appearance, in which the ragged outer spore wall was lost to a large extent. Furthermore, mde10Delta spores were found to be less tolerant to ethanol and diethyl ether than were wild-type spores. The mutagenic replacement of the conserved glutamic acid in the putative protease active site with an alanine residue did not affect the surface morphology or the resistance of spores to environmental stress. Our observations indicate that Mde10 is important in the development of the spore envelope, although this function of Mde10 seems to be independent of its metalloprotease activity.
|Alanine / chemistry Alleles Amino Acid Sequence Base Sequence Binding Sites Blotting, Northern Blotting, Southern Blotting, Western Catalytic Domain Cell Membrane / metabolism Centrifugation, Density Gradient Cysteine / chemistry DNA, Complementary / metabolism Endoplasmic Reticulum / metabolism Epitopes Ethanol / pharmacology Ether / pharmacology Fungal Proteins / metabolism Fungal Proteins / physiology* Gene Deletion Gene Library Genotype Green Fluorescent Proteins Luminescent Proteins / metabolism Meiosis Metalloproteases / chemistry* Metalloproteases / physiology* Microscopy, Electron Microscopy, Fluorescence Models, Biological Molecular Sequence Data Mutagenesis, Site-Directed Mutation Open Reading Frames Plasmids / metabolism Protein Structure, Tertiary Recombinant Fusion Proteins / metabolism Schizosaccharomyces / physiology* Schizosaccharomyces pombe Proteins / chemistry* Schizosaccharomyces pombe Proteins / physiology* Time Factors Transcription, Genetic
|TN8 C996-11D ? FY7539(C525(C)-1A) FY6936(JY878) FY14310(NT10-2) FY14311(NT10-3) FY14312(NT10-9) FY14313(NT10-10) FY14314(NT10-11) FY7051(JZ670) FY7476(AB4)