Dynein Heavy Chain, Encoded by Two Genes in Agaricomycetes, Is Required for Nuclear Migration in Schizophyllum commune

Abstract

The white-rot fungus Schizophyllum commune (Agaricomycetes) was used to study the cell biology of microtubular trafficking during mating interactions, when the two partners exchange nuclei, which are transported along microtubule tracks. For this transport activity, the motor protein dynein is required. In S. commune, the dynein heavy chain is encoded in two parts by two separate genes, dhc1 and dhc2. The N-terminal protein Dhc1 supplies the dimerization domain, while Dhc2 encodes the motor machinery and the microtubule binding domain. This split motor protein is unique to Basidiomycota, where three different sequence patterns suggest independent split events during evolution. To investigate the function of the dynein heavy chain, the gene dhc1 and the motor domain in dhc2 were deleted. Both resulting mutants were viable, but revealed phenotypes in hyphal growth morphology and mating behavior as well as in sexual development. Viability of strain Δdhc2 is due to the higher expression of kinesin-2 and kinesin-14, which was proven via RNA sequencing.

Nuclear movement in fungi

“…1C; Movies 3 and 4) [7, 8]. The role of dynein in positioning nuclei/spindles has subsequently been found in other fungal organisms including Nectria haematococca , Ustilago maydis , Ashbya gossypii , Aspergillus oryzae , Candida albicans and Schizophyllum commune [16–23]. Interestingly, in the dynein-null mutant of A. gossypii , multiple nuclei form a cluster at the hyphal tip [16] (Fig.…”

“…Nuclear movements are important for mating and the subsequent nuclear fusion (Karyogamy) in fungal organisms, and minus-end-directed microtubule motors such as dynein and kinesin-14s are involved in these processes [23, 25, 116–118]. In some filamentous fungi, such as N. crassa and the plant pathogen Fusarium oxysporum , fusion of vegetative hyphae (termed “anastomosis”) occurs during colony formation [119], and hyphal fusion is often followed by nuclear migration, which may be important for colony formation and/or fungal infection [120, 121].…”

Smelling the difference: Transcriptome, proteome and volatilome changes after mating

“…the number of regulated genes and especially the fold changes, are different. The data were validated by qRT-PCR (as described by Brunsch et al 2015, see also Tab. S12).…”

“…2.4 RNA sequencing and microarray analyses RNA was extracted as previously described (Erdmann et al, 2012) and sequenced from mRNA based cDNA libraries by LGC Genomics (Berlin, Germany) as previously described (Brunsch et al, 2015). The data are available at NCBI GEO expression omnibus with the accession number GSE84124.…”

Crosstalk between Ras and inositol phosphate signaling revealed by lithium action on inositol monophosphatase in Schizophyllum commune

“…Differential spots were excised, eluted (Brunsch et al, 2015) and protein identification was performed using MALDI TOF/MS after tryptic digestion (Shevchenko et al, 1996) using 1 μl HCCA matrix (10 mg/ml alpha cyano-4-hydroxy-cinnamic acid in 30% acetonitrile, 70% distilled water with 0.1% trifluoroacetic acid) transferred to an Anchor Chip Target (Bruker Daltonics, Bremen) for 1 μl of the tryptically digested sample. The dried samples were analyzed (ultrafleXtreme MALDI-TOF/TOF; Bruker, Bremen, Germany) and measured using flexControl software followed by analysis with flexAnalysis.…”