Chromosomal loci of Neurospora crassa

Perkins, David; Radford, Alan; Newmeyer, Dorothy; Björkman, Mari

doi:10.1128/mr.46.4.426-570.1982

Cited by 259 publications

(8 citation statements)

References 935 publications

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“…Hygromycin B (Calbiochem) was used at 200 μg/ml to select for the hph gene (Staben et al ., 1989). Scoring for the following N.crassa genes was as described previously: wc‐1 , un‐10 and arg‐10 (Perkins et al ., 1982); am (Kinsey et al ., 1980); and mtr (Irelan et al ., 1994).…”

Section: Methodsmentioning

confidence: 99%

dim-2 encodes a DNA methyltransferase responsible for all known cytosine methylation in Neurospora

2001

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To understand better the control of DNA methylation, we cloned and characterized the dim-2 gene of Neurospora crassa, the only eukaryotic gene currently known in which mutations appear to eliminate DNA methylation. The dim-2 gene is responsible for methylation in both symmetrical and asymmetrical sites. We mapped dim-2 between wc-1 and un-10 on linkage group (LG) VIIR and identi®ed the gene by RFLP mapping and genetic complementation. Dim-2 encodes a 1454 amino acid protein including a C-terminal domain homologous to known DNA methyltransferases (MTases) and a novel N-terminal domain. Neither a deletion that removed the ®rst 186 amino acids of the protein nor a mutation in a putative nucleotide binding site abolished function, but a single amino acid substitution in the predicted catalytic site did. Tests for repeat-induced point mutation (RIP) indicated that dim-2 does not play a role in this process, i.e. duplicated sequences are mutated in dim-2 strains, as usual, but the mutated sequences are not methylated, unlike the situation in dim-2 + strains. We conclude that dim-2 encodes an MTase that is responsible for all DNA methylation in vegetative tissues of Neurospora.

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Section: Methodsmentioning

confidence: 99%

dim-2 encodes a DNA methyltransferase responsible for all known cytosine methylation in Neurospora

2001

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“…To identify the partner cyclin of PHO85-1 involved in the glycogen metabolism control in N. crassa , a BLASTp search in the N. crassa genome database 6 using as queries the S. cerevisiae cyclins, retrieved a homolog protein encoded by the ORF NCU08772, which shares an average of 24% identity to the yeast cyclins mainly in the cyclin domain ( Figure 1A ). The predicted protein is annotated in the fungus database as nuclear division-60, and is named here as PCL-1 consistent with the N. crassa nomenclature ( Perkins et al, 1982 ) and considering that it is the first PCL described in N. crassa . PCL-1 is a non-essential protein, and the Δ pcl-1 mutant strain exhibits wild-type phenotypes suggesting that it is not involved in processes related to growth and development (results not shown).…”

Section: Resultsmentioning

confidence: 99%

“…This protein is annotated in the fungus database as a cyclin-dependent protein kinase, and was formerly named as PGOV ( Peleg et al, 1996 ). Based on its high identity to the S. cerevisiae Pho85 protein, we renamed here as PHO85-1, according to N. crassa nomenclature ( Perkins et al, 1982 ). As previously mentioned, Pho85 CDK requires different cyclins for activation, and, in yeast, the Pcl6, 7, 8, and 10 cyclins are described as partners proteins that target the kinase to glycogen substrate.…”

Section: Introductionmentioning

confidence: 99%

The Neurospora crassa PCL-1 cyclin is a PHO85-1 (PGOV) kinase partner that directs the complex to glycogen metabolism and is involved in calcium metabolism regulation

et al. 2022

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Cyclins are a family of proteins characterized by possessing a cyclin box domain that mediates binding to cyclin dependent kinases (CDKs) partners. In this study, the search for a partner cyclin of the PHO85-1 CDK retrieved PCL-1 an ortholog of yeast Pcls (for Pho85 cyclins) that performs functions common to Pcls belonging to different cyclin families. We show here that PCL-1, as a typical cyclin, is involved in cell cycle control and cell progression. In addition, PCL-1 regulates glycogen metabolism; Δpcl-1 cells accumulate higher glycogen levels than wild-type cells and the glycogen synthase (GSN) enzyme is less phosphorylated and, therefore, more active in the mutant cells. Together with PHO85-1, PCL-1 phosphorylates in vitro GSN at the Ser636 amino acid residue. Modeling studies identified PHO85-1 and PCL-1 as a CDK/cyclin complex, with a conserved intermolecular region stabilized by hydrophobic and polar interactions. PCL-1 is also involved in calcium and NaCl stress response. Δpcl-1 cells are sensitive to high NaCl concentration; on the contrary, they grow better and overexpress calcium responsive genes under high calcium chloride concentration compared to the wild-type strain. The expression of the calcium-responsive CRZ-1 transcription factor is modulated by PCL-1, and this transcription factor seems to be less phosphorylated in Δpcl-1 cells since exhibits nuclear location in these cells in the absence of calcium. Our results show that PCL-1 locates at different cell regions suggesting that it may determine its activity by controlling its intracellular location and reveal an interesting functional divergence between yeast and filamentous fungus cyclins.

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“…The high availability of exogenous tryptophan can also promote feedback repression of the initial steps of aromatic amino acids biosynthesis, and impair trp - mutants’ growth due to decreased synthesis of phenylalanine and tyrosine. To circumvent this defect, simultaneous tryptophan and phenylalanine supply improved the growth of N. crassa trp mutants [ 85 ]. Further investigation can clarify whether CpcA plays the same role in F. pedrosoi and whether the addition of external sources of aromatic amino acids would benefit mycelial growth.…”

Section: Discussionmentioning

confidence: 99%

Expanding the Toolbox for Functional Genomics in Fonsecaea pedrosoi: The Use of Split-Marker and Biolistic Transformation for Inactivation of Tryptophan Synthase (trpB) Gene

Favilla

Herman

Goersch

et al. 2023

JoF

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Chromoblastomycosis (CBM) is a disease caused by several dematiaceous fungi from different genera, and Fonsecaea is the most common which has been clinically isolated. Genetic transformation methods have recently been described; however, molecular tools for the functional study of genes have been scarcely reported for those fungi. In this work, we demonstrated that gene deletion and generation of the null mutant by homologous recombination are achievable for Fonsecaea pedrosoi by the use of two approaches: use of double-joint PCR for cassette construction, followed by delivery of the split-marker by biolistic transformation. Through in silico analyses, we identified that F. pedrosoi presents the complete enzymatic apparatus required for tryptophan (trp) biosynthesis. The gene encoding a tryptophan synthase trpB —which converts chorismate to trp—was disrupted. The ΔtrpB auxotrophic mutant can grow with external trp supply, but germination, viability of conidia, and radial growth are defective compared to the wild-type and reconstituted strains. The use of 5-FAA for selection of trp- phenotypes and for counter-selection of strains carrying the trp gene was also demonstrated. The molecular tools for the functional study of genes, allied to the genetic information from genomic databases, significantly boost our understanding of the biology and pathogenicity of CBM causative agents.

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Chromosomal loci of Neurospora crassa

Cited by 259 publications

References 935 publications

dim-2 encodes a DNA methyltransferase responsible for all known cytosine methylation in Neurospora

dim-2 encodes a DNA methyltransferase responsible for all known cytosine methylation in Neurospora

The Neurospora crassa PCL-1 cyclin is a PHO85-1 (PGOV) kinase partner that directs the complex to glycogen metabolism and is involved in calcium metabolism regulation

Expanding the Toolbox for Functional Genomics in Fonsecaea pedrosoi: The Use of Split-Marker and Biolistic Transformation for Inactivation of Tryptophan Synthase (trpB) Gene

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