Structure of the Aspergillus niger pelA gene and its expression in Aspergillus niger and Aspergillus nidulans

Someren, M.A. Kusters-van; Harmsen, J.A.M.; Kester, H.C.M.; Visser, Jaap

doi:10.1007/bf00318518

Cited by 160 publications

(106 citation statements)

References 20 publications

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“…The mpdA sequence was determined from both strands. Transformation of A. niger was performed as described by Kusters-van Someren et al (24). For cotransformation, the pyrA gene (15) was used as a selective marker.…”

Section: Methodsmentioning

confidence: 99%

Mannitol Is Required for Stress Tolerance inAspergillus nigerConidiospores

et al. 2003

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D-Mannitol is the predominant carbon compound in conidiospores of the filamentous fungus Aspergillus niger and makes up 10 to 15% of the dry weight. A number of physiological functions have been ascribed to mannitol, including serving as a reserve carbon source, as an antioxidant, and to store reducing power. In this study, we cloned and characterized the A. niger mpdA gene, which encodes mannitol 1-phosphate dehydrogenase (MPD), the first enzyme in the mannitol biosynthesis pathway. The mpdA promoter contains putative binding sites for the development-specific transcription factors BRLA and ABAA. Furthermore, increased expression of mpdA in sporulating mycelium suggests that mannitol biosynthesis is, to a certain extent, developmentally regulated in A. niger. Inactivation of mpdA abolished mannitol biosynthesis in growing mycelium and reduced the mannitol level in conidiospores to 30% that in the wild type, indicating that MPD and mannitol 1-phosphate phosphatase form the major metabolic pathway for mannitol biosynthesis in A. niger. The viability of spores after prolonged storage and germination kinetics were normal in an mpdA null mutant, indicating that mannitol does not play an essential role as a reserve carbon source in A. niger conidia. However, conidiospores of a ⌬mpdA strain were extremely sensitive to a variety of stress conditions, including high temperature, oxidative stress and, to a lesser extent, freezing and lyophilization. Since mannitol supplied in the medium during sporulation repaired this deficiency, mannitol appears to be essential for the protection of A. niger spores against cell damage under these stress conditions. Polyols or polyhydroxyalcohols are present in all organisms, from bacteria to animals. In particular, plants and fungi are known to accumulate high levels of polyols intracellularly, up to several hundred millimoles per liter. The filamentous fungus Aspergillus niger produces a number of different polyols, including glycerol, erythritol, and D-mannitol (41). The intracellular concentrations of the individual polyols in A. niger depend on growth conditions and developmental stage, suggesting that polyols have important functions in fungal physiology.The hexitol D-mannitol is accumulated by vascular plants (37) and many fungal species (21). In A. niger conidiospores, D-mannitol is the predominant carbon-containing compound and makes up 10 to 15% of the dry weight (41). The metabolic pathway for mannitol biosynthesis in ascomycete and deuteromycete fungi most likely consists of two steps (19) (Fig. 1). Fructose 6-phosphate is reduced to mannitol 1-phosphate by NAD ϩ -dependent mannitol 1-phosphate dehydrogenase (MPD), followed by the dephosphorylation of mannitol 1-phosphate, which is catalyzed by mannitol 1-phosphate phosphatase (MPP), yielding mannitol. The presence of MPD and MPP in several fungi, such as A. niger (22), Aspergillus nidulans (34), and Alternaria alternata (18), has been demonstrated, but these enzymes are also responsible for mannitol biosynthesis in protozoa...

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Section: Methodsmentioning

confidence: 99%

Mannitol Is Required for Stress Tolerance inAspergillus nigerConidiospores

et al. 2003

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“…Transformation of A. niger was performed as described (Kusters-van Someren et al 1991;Meyer et al 2010). Protoplasts of A. niger were generated according to de Bekker et al (2009).…”

Section: Transformationmentioning

confidence: 99%

FluG affects secretion in colonies of Aspergillus niger

Wang

Krijgsheld

Hulsman

et al. 2014

Antonie van Leeuwenhoek

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Colonies of Aspergillus niger are characterized by zonal heterogeneity in growth, sporulation, gene expression and secretion. For instance, the glucoamylase gene glaA is more highly expressed at the periphery of colonies when compared to the center. As a consequence, its encoded protein GlaA is mainly secreted at the outer part of the colony. Here, multiple copies of amyR were introduced in A. niger. Most transformants over-expressing this regulatory gene of amylolytic genes still displayed heterogeneous glaA expression and GlaA secretion. However, heterogeneity was abolished in transformant UU-A001.13 by expressing glaA and secreting GlaA throughout the mycelium. Sequencing the genome of UU-A001.13 revealed that transformation had been accompanied by deletion of part of the fluG gene and disrupting its 3 0 end by integration of a transformation vector. Inactivation of fluG in the wild-type background of A. niger also resulted in breakdown of starch under the whole colony. Asexual development of the DfluG strain was not affected, unlike what was previously shown in Aspergillus nidulans. Genes encoding proteins with a signal sequence for secretion, including part of the amylolytic genes, were more often downregulated in the central zone of maltose-grown DfluG colonies and upregulated in the intermediate part and periphery when compared to the wild-type. Together, these data indicate that FluG of A. niger is a repressor of secretion.Keywords Fungus Á Conidiogenesis Á Asexual development Á Aspergillus fluG Á Secretion Á Heterogeneity Electronic supplementary material The online version of this article

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“…Protoplasts were prepared after harvesting the mycelium by treatment with 10 µg/ml Caylase C4 (Cayla, Toulouse, France) and transformed as described by Kusters-van Someren et al [21]. Prior to imaging, coverslips were inoculated with 100 µl of the A. niger spore suspension and incubated for 16 hours at 30 ºC.…”

Section: Accepted Manuscriptmentioning

confidence: 99%

High-precision FLIM–FRET in fixed and living cells reveals heterogeneity in a simple CFP–YFP fusion protein

Millington

Grindlay

Altenbach

et al. 2007

Biophysical Chemistry

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This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. A C C E P T E D M A N U S C R I P T ACCEPTED MANUSCRIPT High-Precision FLIM-FRET in fixed and living cells reveals heterogeneity in a simple CFP-YFP fusion proteinMichael Millington, [a, b] G. Joan Grindlay, [c] Kirsten Altenbach, [a, d] Robert K. Neely, [a, b] Walter Kolch, [ c, e] Mojca Bencina , [ f ] Nick D. Read, [a, d] Anita C. Jones, [a, b] David T. F.Dryden, [a, b]

show abstract

Structure of the Aspergillus niger pelA gene and its expression in Aspergillus niger and Aspergillus nidulans

Cited by 160 publications

References 20 publications

Mannitol Is Required for Stress Tolerance inAspergillus nigerConidiospores

Mannitol Is Required for Stress Tolerance inAspergillus nigerConidiospores

FluG affects secretion in colonies of Aspergillus niger

High-precision FLIM–FRET in fixed and living cells reveals heterogeneity in a simple CFP–YFP fusion protein

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