Dynamin-like protein-dependent formation of Woronin bodies in Saccharomyces cerevisiae upon heterologous expression of a single protein.
Filamentous ascomycetes harbor Woronin bodies and glyoxysomes, two types of microbodies, within one cell at the same time. The dominant protein of the Neurospora crassa Woronin body, HEX1, forms a hexagonal core crystal via oligomerization and evidence has accumulated that Woronin bodies bud off from glyoxysomes. We analyzed whether HEX1 is sufficient to induce Woronin body formation upon heterologous expression in Saccharomyces cerevisiae, an organism devoid of this specialized organelle. In wild-type strain BY4742, initial import of HEX1 into existing peroxisomes enabled the formation of organelles with a hexagonal crystal. The observed structures mimicked the shape of genuine Woronin bodies, but exhibited a lower density and were significantly larger. Double-immunofluorescence analysis revealed that hexagonal HEX1 structures only occasionally co-localized with peroxisomal marker proteins, indicating that the Woronin-body-like structures are well separated from peroxisomes. In cells lacking Vps1p and Dnm1p, dynamin-like proteins required for the division of peroxisomes, the Woronin-body-like organelles remained attached to peroxisomes. The data indicate that Woronin bodies emerge after the formation of a HEX1 core crystal within peroxisomes followed by Vps1p- and Dnm1p-mediated fission.
Würtz
Christian
C
Institut für Physiologische Chemie, Ruhr-Universität Bochum, Germany.
Schliebs
Wolfgang
W
Erdmann
Ralf
R
Rottensteiner
Hanspeter
H
D003460:Crystallization; D034281:Dynamins / Q000378:metabolism*; D005090:Exodeoxyribonucleases / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D008856:Microscopy, Fluorescence; D024101:Mitochondrial Proteins; D008954:Models, Biological; D009492:Neurospora crassa; D015388:Organelles / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D033921:Vesicular Transport Proteins
D005656:Fungal Proteins; D024101:Mitochondrial Proteins; D029701:Saccharomyces cerevisiae Proteins; D033921:Vesicular Transport Proteins; D005090:Exodeoxyribonucleases; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins; C064553:VPS1 protein, S cerevisiae; C094774:DNM1 protein, S cerevisiae; D034281:Dynamins