Developmental expression of genes involved in conidiation and amino acid biosynthesis in Neurospora crassa

Sachs, Matthew S.; Yanofsky, Charles

doi:10.1016/0012-1606(91)90322-t

Cited by 104 publications

(112 citation statements)

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“…Here, we show that prnB message accumulates during conidiospore germination and that this accumulation is partially independent of proline induction and/or of a functional prnA gene product. It is known from studies with A. nidulans and N. crassa that amino acid pools, and especially proline pools, reduce dramatically upon germination (Van Etten et al, 1983;Cook and Anthony, 1978;Sachs and Yanofsky, 1991). Moreover, it has also been shown that transcription of amino acid biosynthetic genes is activated specifically during germination of N. crassa conidia .…”

Section: Discussionmentioning

confidence: 99%

“…The analogue 3-amino-1,2,4-triazole (3AT) has been shown to lead to histidine starvation through inhibition of histidine biosynthesis both in S. cerevisiae and Neurospora crassa (Hinnebusch, 1988;Sachs and Yanofsky, 1991;Sachs, 1996). We compared the induction of prnB, prnC, and prnD by proline and by 3AT in mycelia of prnA þ and prnA ¹ strains (Fig.…”

Section: Proline Transport Is Activated During Conidiospore Germinationmentioning

confidence: 99%

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The gene encoding the major proline transporter of Aspergillus nidulans is upregulated during conidiospore germination and in response to proline induction and amino acid starvation

Tazebay

Sophianopoulou

Scazzocchio

et al. 1997

Molecular Microbiology

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SummaryIn Aspergillus nidulans a highly specific L-proline transporter is encoded by the prnB gene which is tightly linked to all other genes involved in proline catabolism. In mycelia, the expression of the prn structural genes is finely co-regulated in response to proline induction and nitrogen/carbon catabolite repression. In this study we establish that prnB expression is also activated during germination of conidiospores. This activation persists until the development of 6 h-old mycelia and it is independent of proline induction mediated by the pathway-specific prnA gene product. We then show that, in mycelia, prnB transcription is activated in response to proline or histidine starvation. This process has two components: a prnA-dependent and a prnA-independent component. A cis-acting element that conforms to the consensus target of the GCN4/CPC1 transcriptional activators mediating amino acid biosynthesis activation in other fungi is involved in the activation of prnB transcription in response to amino acid starvation. We also show that the stimulation of prnB expression in germinating conidiospores is not due exclusively to transient internal amino acid starvation occurring during the transition from conidiospore to mycelium. This is the first report that an amino acid transporter gene is upregulated during development and in response to amino acid starvation and specific amino acid induction.

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Section: Discussionmentioning

confidence: 99%

Section: Proline Transport Is Activated During Conidiospore Germinationmentioning

confidence: 99%

The gene encoding the major proline transporter of Aspergillus nidulans is upregulated during conidiospore germination and in response to proline induction and amino acid starvation

Tazebay

Sophianopoulou

Scazzocchio

et al. 1997

Molecular Microbiology

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“…Samples containing 1-5 x 106 irradiated conidia were suspended in 15 ml of Vogel's minimal agar plus sorbose (18,19) plus hygromycin at 1.5 mg/ml and poured into a 100-mm Petri dish. After the agar in each plate solidified, 20 (18).…”

Section: Methodsmentioning

confidence: 99%

“…RNA was isolated from mycelia or conidia by disrupting suspensions with 0.5-mm glass beads in a mini-beadbeater (Biospec Products, Bartlesville, OK) in the presence of phenol/chloroform and SDScontaining buffer, as described (20), except that 10 mM EDTA was added to the extraction buffer. Radiolabeled RNA probes complementary to con-6, con-8, con-10, and eas mRNAs were prepared by in vitro transcription from linearized subclones prepared in this laboratory-pCON6-6, pRB2, pBW100, and pEAS, respectively-using either 17 or T3 RNA polymerases.…”

Section: Methodsmentioning

confidence: 99%

Mutants of Neurospora crassa that alter gene expression and conidia development.

Madi

Ebbole²,

White³

et al. 1994

Proc. Natl. Acad. Sci. U.S.A.

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Several genes have been identied that are highly expressed during conidiation. Inactivation ofthese genes has no observable phenotypic effect. Transcripts of two such genes, con-6 and con-10, are norally absent from vegetative mycella. To identify regulatory genes that affect con-6 and/or con-10 expression, strains were prepared in which the regulatory regions for these genes were fused to a gene conferrin hygromycin resistance. Mutants were then selected that were resistant to the drug during mycelial growth. Mutations in several of the isolates had trans effects; they activated transcription of the corresponding intact gene and, in most oates, one or more ofthe other con genes. Most Analyses of the protein and mRNA species present in mycelia, aerial hyphae, and conidia have revealed significant differences in gene expression (3)(4)(5). A differential screening procedure was used to clone genes that are preferentially expressed during conidiation (4). Four of these genes, con-6, con-8, con-10, and con-13, have been sequenced, and their expression has been examined (6-10). A fifth conidiationspecific gene, eas, also has been characterized (11, 12); eas encodes the rodlet protein that coats the surface ofthe mature conidium.con-6 and con-10 encode 93-and 86-residue polypeptides, respectively; the functions of these polypeptides are unknown. con-6 and con-10 transcripts and proteins are normally undetectable in vegetative mycelium but appear at high levels during conidiation and in mature conidia; these transcripts and proteins disappear a few hours after conidia germinate (6,13). The expression pattern of these genes closely parallels that of eas (11, 12). The con-6 and con-10 proteins are present in a second type of asexual spore, the microconidium (10,13), and the con-10 protein is present in sexual spores (10, 13). Expression of con-6 and con-10 can also be induced in vegetative mycelial cultures by light exposure or by imposing conditions that induce circadian rhythm (10,(13)(14)(15). We describe here the selection and partial characterization of mutants that express con-6 and con-10 aberrantly during vegetative mycelial growth. Some of these mutants were found to be blocked in conidiation, whereas others exhibited different developmental abnormalities. MATERIALS AND METHODSStrains and Pids. Strain CH10 was constructed by homologous integration, using pCH10, a plasmid containing a con-10 translational fusion to the hygromycin phosphotransferase gene (hph) (16). Integration was at the his-3 locus of Fungal Genetics Stock Center (FGSC) strain 462: his-3 A. The integrated plasmid consisted of pBluescript KS+ (Promega) vector containing con-10 DNA from positions 1139 to 1935 (8); this segment contains the first 40 codons of con-10. The con-10 DNA is fused in-frame to the second codon of hph followed by the Aspergillus nidulans trpC transcription termination region from plasmid pDH25 (16). The plasmid also contains a 5' truncated copy of the his-3 gene that allows reconstitution of an intact copy of the ...

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“…VM plates were grown in the dark at 30°C for 3 d, whereas SCM plates were grown in constant light at 25°C for 6 d. Submerged vegetative cultures were obtained by inoculation of liquid VM with 5-8-d old macroconidia to a final concentration of 1 ϫ 10 6 macroconidia/ml followed by culturing at 30°C for 16 h with shaking at 200 rpm, whereas 3-d-old liquid SCM cultures were grown with constant light at room temperature at 60 rpm. Total RNA was extracted as described previously (Sachs and Yanofsky, 1991) or with the TRIzol reagent according to the manufacturer's recommendations (Invitrogen, Carlsbad, CA) or by using the PureScript kit according to the manufacturer's directions for plant tissue (Gentra Systems, Minneapolis, MN). …”

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confidence: 99%

The Response Regulator RRG-1 Functions Upstream of a Mitogen-activated Protein Kinase Pathway Impacting Asexual Development, Female Fertility, Osmotic Stress, and Fungicide Resistance inNeurospora crassa

Jones

Greer‐Phillips²,

Borkovich

2007

MBoC

105

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Two-component systems, consisting of proteins with histidine kinase and/or response regulator domains, regulate environmental responses in bacteria, Archaea, fungi, slime molds, and plants. Here, we characterize RRG-1, a response regulator protein from the filamentous fungus Neurospora crassa. The cell lysis phenotype of ⌬rrg-1 mutants is reminiscent of osmotic-sensitive (os) mutants, including nik-1/os-1 (a histidine kinase) and strains defective in components of a mitogen-activated protein kinase (MAPK) pathway: os-4 (MAPK kinase kinase), os-5 (MAPK kinase), and os-2 (MAPK). Similar to os mutants, ⌬rrg-1 strains are sensitive to hyperosmotic conditions, and they are resistant to the fungicides fludioxonil and iprodione. Like os-5, os-4, and os-2 mutants, but in contrast to nik-1/os-1 strains, ⌬rrg-1 mutants do not produce female reproductive structures (protoperithecia) when nitrogen starved. OS-2-phosphate levels are elevated in wild-type cells exposed to NaCl or fludioxonil, but they are nearly undetectable in ⌬rrg-1 strains. OS-2-phosphate levels are also low in ⌬rrg-1, os-2, and os-4 mutants under nitrogen starvation. Analysis of the rrg-1 D921N allele, mutated in the predicted phosphorylation site, provides support for phosphorylation-dependent and -independent functions for RRG-1. The data indicate that RRG-1 controls vegetative cell integrity, hyperosmotic sensitivity, fungicide resistance, and protoperithecial development through regulation of the OS-4/OS-5/OS-2 MAPK pathway. INTRODUCTIONTwo-component regulatory systems are signal transduction pathways found in bacteria, Archaea, slime molds, fungi, and plants (for reviews, see Wolanin et al., 2002). Twocomponent systems have been implicated in responses to light, osmolarity, cellular redox status, nutrient and oxygen levels, virulence, and other factors (for review, see Wolanin et al., 2002;Borkovich et al., 2004). A basic two-component system consists of a histidine kinase and a response regulator (for review, see West and Stock, 2001;Wolanin et al., 2002). Histidine kinases autophosphorylate on a conserved histidine residue in the histidine kinase domain. The phosphoryl group is then transferred to an aspartate residue in the receiver domain on the response regulator. Complex two-component signaling pathways (also termed phosphorelays) contain hybrid histidine kinases that have both a histidine kinase domain and a response regulator receiver domain in the same protein, thus facilitating an intramolecular phosphate transfer reaction. The phosphate group on the aspartate residue is transferred to a histidine phosphotransfer (HPT) protein. Subsequently, the histidine phosphotransfer protein phosphorylates the receiver domain on a response regulator. Phosphorelays are the major two-component pathways in eukaryotic systems; however, two-component systems are not present in animals, making them ideal candidate targets for antimicrobial drugs (for review, see Alex et al., 1998;Wolanin et al., 2002;Borkovich et al., 2004).Many response regulators contain a ...

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Developmental expression of genes involved in conidiation and amino acid biosynthesis in Neurospora crassa

Cited by 104 publications

References 49 publications

The gene encoding the major proline transporter of Aspergillus nidulans is upregulated during conidiospore germination and in response to proline induction and amino acid starvation

The gene encoding the major proline transporter of Aspergillus nidulans is upregulated during conidiospore germination and in response to proline induction and amino acid starvation

Mutants of Neurospora crassa that alter gene expression and conidia development.

The Response Regulator RRG-1 Functions Upstream of a Mitogen-activated Protein Kinase Pathway Impacting Asexual Development, Female Fertility, Osmotic Stress, and Fungicide Resistance inNeurospora crassa

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