On the Evidence Supporting That AN11127 Encodes an Aspergillus Nidulans Sec12 Orthologous Protein. Reply to Bravo-Plaza et al. Comment on “Dimou et al. Profile of Membrane Cargo Trafficking Proteins and Transporters Expressed under N Source Derepressing Conditions in Aspergillus nidulans. J. Fungi 2021, 7, 560”

Recent biochemical and biophysical evidence have established that membrane lipids, namely phospholipids, sphingolipids and sterols, are critical for the function of eukaryotic plasma membrane transporters. Here, we study the effect of selected membrane lipid biosynthesis mutations and of the ergosterol-related antifungal itraconazole on the subcellular localization, stability and transport kinetics of two well-studied purine transporters, UapA and AzgA, in Aspergillus nidulans. We show that genetic reduction in biosynthesis of ergosterol, sphingolipids or phosphoinositides arrest A. nidulans growth after germling formation, but solely blocks in early steps of ergosterol (Erg11) or sphingolipid (BasA) synthesis have a negative effect on plasma membrane (PM) localization and stability of transporters before growth arrest. Surprisingly, the fraction of UapA or AzgA that reaches the PM in lipid biosynthesis mutants is shown to conserve normal apparent transport kinetics. We further show that turnover of UapA, which is the transporter mostly sensitive to membrane lipid content modification, occurs during its trafficking and by enhanced endocytosis, and is partly dependent on autophagy and Hect-type HulARsp5 ubiquitination. Our results point out that the role of specific membrane lipids on transporter biogenesis and function in vivo is complex, combinatorial and transporter-dependent.

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The splicing-factor Prp40 affects dynein–dynactin function in Aspergillus nidulans

Molecular Biology of the Cell ◽

10.1091/mbc.e20-03-0166 ◽

2020 ◽

Vol 31 (12) ◽

pp. 1289-1301 ◽

Cited By ~ 1

Author(s):

Rongde Qiu ◽

Jun Zhang ◽

Xin Xiang

Keyword(s):

Aspergillus Nidulans ◽

Early Endosome ◽

Splicing Factor ◽

Factors Affecting ◽

Mutant Screen ◽

And Function

We performed a genomewide mutant screen for genes affecting dynein-mediated early-endosome distribution in Aspergillus nidulans, and we unexpectedly identified Prp40A, a homologue of the yeast splicing factor Prp40. Prp40A and its higher eukaryotic homologues may represent new factors affecting the assembly and function of dynein-dynactin.

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Suppression of the bimC4 mitotic spindle defect by deletion of klpA, a gene encoding a KAR3-related kinesin-like protein in Aspergillus nidulans.

The Journal of Cell Biology ◽

10.1083/jcb.120.1.153 ◽

1993 ◽

Vol 120 (1) ◽

pp. 153-162 ◽

Cited By ~ 90

Author(s):

M J O'Connell ◽

P B Meluh ◽

M D Rose ◽

N R Morris

Keyword(s):

Aspergillus Nidulans ◽

Mitotic Spindle ◽

Structural Similarity ◽

Temperature Sensitive ◽

Gene Encoding ◽

Expression Studies ◽

Primary Amino ◽

And Function ◽

Spindle Function ◽

The Relationship

To investigate the relationship between structure and function of kinesin-like proteins, we have identified by polymerase chain reaction (PCR) a new kinesin-like protein in the filamentous fungus Aspergillus nidulans, which we have designated KLPA. DNA sequence analysis showed that the predicted KLPA protein contains a COOH terminal kinesin-like motor domain. Despite the structural similarity of KLPA to the KAR3 and NCD kinesin-like proteins of Saccharomyces cerevisiae and Drosophila melanogaster, which also posses COOH-terminal kinesin-like motor domains, there are no significant sequence similarities between the nonmotor or tail portions of these proteins. Nevertheless, expression studies in S. cerevisiae showed that klpA can complement a null mutation in KAR3, indicating that primary amino acid sequence conservation between the tail domains of kinesin-like proteins is not necessarily required for conserved function. Chromosomal deletion of the klpA gene exerted no observable mutant phenotype, suggesting that in A. nidulans there are likely to be other proteins functionally redundant with KLPA. Interestingly, the temperature sensitive phenotype of a mutation in another gene, bimC, which encodes a kinesin-like protein involved in mitotic spindle function in A. nidulans, was suppressed by deletion of klpA. We hypothesize that the loss of KLPA function redresses unbalanced forces within the spindle caused by mutation in bimC, and that the KLPA and BIMC kinesin-like proteins may play opposing roles in spindle function.

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Gene Structure and Function of fkhE, a Forkhead Gene in a Filamentous Fungus Aspergillus nidulans

The Korean Journal of Mycology ◽

10.4489/kjm.2010.38.2.160 ◽

2010 ◽

Vol 38 (2) ◽

pp. 160-166 ◽

Cited By ~ 3

Author(s):

Mi-Hye Park ◽

Hyoun-Young Kim ◽

Jong-Hwa Kim ◽

Kap-Hoon Han

Keyword(s):

Aspergillus Nidulans ◽

Gene Structure ◽

Filamentous Fungus ◽

Structure And Function ◽

And Function ◽

Forkhead Gene

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Conservation of structure and function of the aflatoxin regulatory gene aflR from Aspergillus nidulans and A. flavus

Current Genetics ◽

10.1007/s002940050084 ◽

1996 ◽

Vol 29 (6) ◽

pp. 549-555 ◽

Cited By ~ 12

Author(s):

J.-H. Yu ◽

Robert A. E. Butchko ◽

Mary Fernandes ◽

N. P. Keller ◽

Thomas J. Leonard ◽

...

Keyword(s):

Aspergillus Nidulans ◽

Regulatory Gene ◽

Structure And Function ◽

And Function

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Improved gene expression in Aspergillus nidulans

Canadian Journal of Botany ◽

10.1139/b95-334 ◽

1995 ◽

Vol 73 (S1) ◽

pp. 876-884 ◽

Cited By ~ 19

Author(s):

William E. Hintz ◽

Inge Kalsner ◽

Ewa Plawinski ◽

Zimin Guo ◽

Peter A. Lagosky

Keyword(s):

Gene Expression ◽

Alcohol Dehydrogenase ◽

Aspergillus Nidulans ◽

Glucose Repression ◽

Expression System ◽

Therapeutic Proteins ◽

Regulatory Sequences ◽

Transcriptional Regulatory ◽

And Function

A variety of gene expression systems have been developed that utilize the promoter and transcriptional regulatory sequences derived from carbon-catabolite repressed genes for the expression of heterologous genes. The alcA expression system of Aspergillus nidulans utilizes the promoter and regulatory sequences derived from the alcohol dehydrogenase I (alcA) gene. Expression of the alcA gene is repressed by a DNA-binding protein (CreA) in the presence of glucose and induced by ethanol under glucose-depleted conditions. One problem encountered during the expression of therapeutic proteins in A. nidulans is the coexpression of secreted proteases at the time of maximal secretion of heterologous product. To avoid the proteases we created an alcA promoter variant that is no longer sensitive to glucose repression hence could drive expression at earlier time points during the fermentation. The use of this promoter variant in the expression of recombinant interleukin-6 is discussed. A second problem encountered during the expression of high-quality human therapeutic proteins in Aspergillus is aberrant glycosylation. Lower eukaryotic systems, such as Aspergillus, tend to add highly branched mannosidic chains to heterologous secreted protein products. N-Glycans can be important for both the structure and function of specific glycoproteins, hence efforts are being made to in vivo alter the type and complexity of N-glycans substituted by A. nidulans. Key words: Aspergillus, gene expression, alcohol dehydrogenase, glycosylation.

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Structure and function of a broad-specificity chitin deacetylase from Aspergillus nidulans FGSC A4

Scientific Reports ◽

10.1038/s41598-017-02043-1 ◽

2017 ◽

Vol 7 (1) ◽

Cited By ~ 27

Author(s):

Zhanliang Liu ◽

Laurie M. Gay ◽

Tina R. Tuveng ◽

Jane W. Agger ◽

Bjørge Westereng ◽

...

Keyword(s):

Aspergillus Nidulans ◽

Structure And Function ◽

Chitin Deacetylase ◽

And Function ◽

Broad Specificity

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Transporter oligomerization: form and function

Biochemical Society Transactions ◽

10.1042/bst20160217 ◽

2016 ◽

Vol 44 (6) ◽

pp. 1737-1744 ◽

Cited By ~ 23

Author(s):

Yilmaz Alguel ◽

Alexander D. Cameron ◽

George Diallinas ◽

Bernadette Byrne

Keyword(s):

Membrane Proteins ◽

Aspergillus Nidulans ◽

Membrane Trafficking ◽

Transport Protein ◽

Integral Membrane Proteins ◽

Structural Basis ◽

Protein Oligomerization ◽

Form And Function ◽

The Individual ◽

And Function

Transporters are integral membrane proteins with central roles in the efficient movement of molecules across biological membranes. Many transporters exist as oligomers in the membrane. Depending on the individual transport protein, oligomerization can have roles in membrane trafficking, function, regulation and turnover. For example, our recent studies on UapA, a nucleobase ascorbate transporter, from Aspergillus nidulans, have revealed both that dimerization of this protein is essential for correct trafficking to the membrane and the structural basis of how one UapA protomer can affect the function of the closely associated adjacent protomer. Here, we review the roles of oligomerization in many particularly well-studied transporters and transporter families.

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The Septin AspB in Aspergillus nidulans Forms Bars and Filaments and Plays Roles in Growth Emergence and Conidiation

Eukaryotic Cell ◽

10.1128/ec.05164-11 ◽

2012 ◽

Vol 11 (3) ◽

pp. 311-323 ◽

Cited By ~ 44

Author(s):

Yainitza Hernández-Rodríguez ◽

Susan Hastings ◽

Michelle Momany

Keyword(s):

Aspergillus Nidulans ◽

Filamentous Fungi ◽

Germ Tube ◽

Fluorescent Protein ◽

Early Growth ◽

Content Type ◽

Green Fluorescent ◽

And Function ◽

Cytoskeletal Elements ◽

Germ Tubes

ABSTRACTIn yeast, septins form rings at the mother-bud neck and function as diffusion barriers. In animals, septins form filaments that can colocalize with other cytoskeletal elements. In the filamentous fungusAspergillus nidulansthere are five septin genes,aspA(an ortholog ofSaccharomyces cerevisiae CDC11),aspB(an ortholog ofS. cerevisiae CDC3),aspC(an ortholog ofS. cerevisiae CDC12),aspD(an ortholog ofS. cerevisiae CDC10), andaspE(found only in filamentous fungi). TheaspBgene was previously reported to be the most highly expressedAspergillus nidulansseptin and to be essential. Using improved gene targeting techniques, we found that deletion ofaspBis not lethal but results in delayed septation, increased emergence of germ tubes and branches, and greatly reduced conidiation. We also found that AspB-green fluorescent protein (GFP) localizes as rings and collars at septa, branches, and emerging layers of the conidiophore and as bars and filaments in conidia and hyphae. Bars are found in dormant and isotropically expanding conidia and in subapical nongrowing regions of hyphae and display fast movements. Filaments form as the germ tube emerges, localize to hyphal and branch tips, and display slower movements. All visible AspB-GFP structures are retained inΔaspDand lost inΔaspAandΔaspCstrains. Interestingly, in theΔaspEmutant, AspB-GFP rings, bars, and filaments are visible in early growth, but AspB-GFP rods and filaments disappear after septum formation. AspE orthologs are only found in filamentous fungi, suggesting that this class of septins might be required for stability of septin bars and filaments in highly polar cells.

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