Disruption of the Ao
            <i>pex11-1</i>
            Gene Involved in Peroxisome Proliferation Leads to Impaired Woronin Body Formation in
            <i>Aspergillus oryzae</i>

Escaño, Cristopher Salazar; Juvvadi, Praveen R.; Jin, Feng; Takahashi, Tadashi; Koyama, Yasuji; Yamashita, Shizuya; Maruyama, Jun‐ichi; Kitamoto, Katsuhiko

doi:10.1128/ec.00197-08

Cited by 56 publications

(68 citation statements)

References 52 publications

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“…The PCR product was digested with SmaI and then inserted into the SmaI site of the plasmid pgEHH (20), and the plasmid pgEH2B-3 was constructed. The generated 5Ј entry clone and center entry clone plasmids (pg5ЈPH-2 and pgEH2B-3), 3Ј entry clone plasmid pg3ЈDRM-CF (mdsred) (9), and the destination vector pgDSO containing the A. oryzae sC gene as a selectable marker (9), were used for Multisite Gateway LR reaction. The generated plasmid was named pgHHDR and introduced into the NS4 (niaD Ϫ sC Ϫ ) strain (37).…”

Section: Methodsmentioning

confidence: 99%

SclR, a Basic Helix-Loop-Helix Transcription Factor, Regulates Hyphal Morphology and Promotes Sclerotial Formation in Aspergillus oryzae

et al. 2011

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Most known basic-region helix-loop-helix (bHLH) proteins belong to a superfamily of transcription factors often involved in the control of growth and differentiation. Therefore, inappropriate expression of genes encoding bHLH proteins is frequently associated with developmental dysfunction. In our previously reported study, a novel bHLH protein-encoding gene (AO090011000215) of Aspergillus oryzae was identified. The genedisrupted strain was found to produce dense conidia, but sparse sclerotia, relative to the parent strain. Here, to further analyze its function, we generated an overexpressing strain using the A. oryzae amyB gene promoter. Genetic overexpression led to a large number of initial hyphal aggregations and then the formation of mature sclerotia; it was therefore designated sclR (sclerotium regulator). At the same time, the sclR-overexpressing strain also displayed both delayed and decreased conidiation. Scanning electron microscopy indicated that the aerial hyphae of the sclR-overexpressing strain were extremely branched and intertwined with each other. In the generation of the SclR-enhanced green fluorescent protein (EGFP) expression strain, the SclR-EGFP protein fusion was conditionally detected in the nuclei. In addition, the loss of sclR function led to rapid protein degradation and cell lysis in dextrin-polypeptone-yeast extract liquid medium. Taken together, these observations indicate that SclR plays an important role in hyphal morphology, asexual conidiospore formation, and the promotion of sclerotial production, even retaining normal cell function, at least in submerged liquid culture.

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Section: Methodsmentioning

confidence: 99%

SclR, a Basic Helix-Loop-Helix Transcription Factor, Regulates Hyphal Morphology and Promotes Sclerotial Formation in Aspergillus oryzae

et al. 2011

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“…In this assay, hyphal tip bursting is induced by flooding A. oryzae colonies with water, and the ability of hyphal cells to prevent the excessive loss of cytoplasm is assessed. This ability is lost in hyphae defective for Woronin body formation (9), and excessive cytoplasmic loss also partially occurs as a result of a deficiency in Woronin body differentiation from peroxisomes (18). Using this quantitative assay, here, the roles of the A. oryzae LAH (AoLAH) protein in the tethering and septal plugging functions of Woronin bodies were investigated.…”

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confidence: 99%

A Large Nonconserved Region of the Tethering Protein Leashin Is Involved in Regulating the Position, Movement, and Function of Woronin Bodies in Aspergillus oryzae

et al. 2014

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The Woronin body is a Pezizomycotina-specific organelle that is typically tethered to the septum, but upon hyphal wounding, it plugs the septal pore to prevent excessive cytoplasmic loss. Leashin (LAH) is a large Woronin body tethering protein that contains highly conserved N-and C-terminal regions and a long (ϳ2,500-amino-acid) nonconserved middle region. As the involvement of the nonconserved region in Woronin body function has not been investigated, here, we functionally characterized individual regions of the LAH protein of Aspergillus oryzae (AoLAH). In an Aolah disruptant, no Woronin bodies were tethered to the septum, and hyphae had a reduced ability to prevent excessive cytoplasmic loss upon hyphal wounding. Localization analysis revealed that the N-terminal region of AoLAH associated with Woronin bodies dependently on AoWSC, which is homologous to Neurospora crassa WSC (Woronin body sorting complex), and that the C-terminal region was localized to the septum. Elastic movement of Woronin bodies was observed when visualized with an AoLAH N-terminal-region-enhanced green fluorescent protein (EGFP) fusion protein. An N-and C-terminal fusion construct lacking the nonconserved middle region of AoLAH was sufficient for the tethering of Woronin bodies to the septum. However, Woronin bodies were located closer to the septum and exhibited impaired elastic movement. Moreover, expression of middle-region-deleted AoLAH in the Aolah disruptant did not restore the ability to prevent excessive cytoplasmic loss. These findings indicate that the nonconserved middle region of AoLAH has functional importance for regulating the position, movement, and function of Woronin bodies.

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“…The Hex1 protein self-assembles to form the Woronin body (WB). The WB is a peroxisome-derived dense-core microbody that is specific to filamentous ascomycetes (21)(22)(23)(24)(25)(26)(27)(28)46). WBs maintain cellular integrity by sealing the septal pore after cellular damage in Neurospora crassa (23)(24)(25).…”

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The HEX1 Gene of Fusarium graminearum Is Required for Fungal Asexual Reproduction and Pathogenesis and for Efficient Viral RNA Accumulation of Fusarium graminearum Virus 1

Son

Lee

et al. 2013

J Virol

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bThe accumulation of viral RNA depends on many host cellular factors. The hexagonal peroxisome (Hex1) protein is a fungal protein that is highly expressed when the DK21 strain of Fusarium graminearum virus 1 (FgV1) infects its host, and Hex1 affects the accumulation of FgV1 RNA. The Hex1 protein is the major constituent of the Woronin body (WB), which is a peroxisome-derived electron-dense core organelle that seals the septal pore in response to hyphal wounding. To clarify the role of Hex1 and the WB in the relationship between FgV1 and Fusarium graminearum, we generated targeted gene deletion and overexpression mutants. Although neither HEX1 gene deletion nor overexpression substantially affected vegetative growth, both changes reduced the production of asexual spores and reduced virulence on wheat spikelets in the absence of FgV1 infection. However, the vegetative growth of deletion and overexpression mutants was increased and decreased, respectively, upon FgV1 infection compared to that of an FgV1-infected wild-type isolate. Viral RNA accumulation was significantly decreased in deletion mutants but was significantly increased in overexpression mutants compared to the viral RNA accumulation in the virus-infected wild-type control. Overall, these data indicate that the HEX1 gene plays a direct role in the asexual reproduction and virulence of F. graminearum and facilitates viral RNA accumulation in the FgV1-infected host fungus.T he interactions between viral elements and host factors are important for maintaining the infection cycles of RNA viruses in host cells. Viruses utilize numerous host factors that play essential roles in virus infection. Therefore, understanding the role(s) of host factors can provide insight into the molecular mechanism(s) of host and virus interactions. Relative to host factors affecting RNA viruses of animals and plants, host factors affecting fungal viruses (mycoviruses) are poorly understood.Several host and viral components required for virus life in cells have been identified and characterized in the model organism Saccharomyces cerevisiae. The knowledge obtained by studying S. cerevisiae as a host for several double-stranded RNA (dsRNA) and single-stranded RNA (ssRNA) viruses has greatly extended our understanding of mycovirus-host interaction (1). With respect to filamentous fungi, the host factors required for mycovirus replication and symptom induction have been well described for the interaction between the prototypic hypovirus Cryphonectria hypovirus 1 strain EP713 (CHV1) and its host, the chestnut blight fungus (Cryphonectria parasitica). One of these host factors, NAM-1, modulates symptom induction in the fungus in response to CHV1 infection (2). The hypovirus-responsive host transcription factor gene pro1 is required for female fertility of C. parasitica, development of its asexual spores, and the maintenance of CHV1 infection (3). The host gene Cpbir1 (bir1 of C. parasitica), which encodes the IAP (inhibitor of apoptosis protein) CpBir1 and which is required for fungal...

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Disruption of the Ao pex11-1 Gene Involved in Peroxisome Proliferation Leads to Impaired Woronin Body Formation in Aspergillus oryzae

Cited by 56 publications

References 52 publications

SclR, a Basic Helix-Loop-Helix Transcription Factor, Regulates Hyphal Morphology and Promotes Sclerotial Formation in Aspergillus oryzae

SclR, a Basic Helix-Loop-Helix Transcription Factor, Regulates Hyphal Morphology and Promotes Sclerotial Formation in Aspergillus oryzae

A Large Nonconserved Region of the Tethering Protein Leashin Is Involved in Regulating the Position, Movement, and Function of Woronin Bodies in Aspergillus oryzae

The HEX1 Gene of Fusarium graminearum Is Required for Fungal Asexual Reproduction and Pathogenesis and for Efficient Viral RNA Accumulation of Fusarium graminearum Virus 1

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