Aspergillus fumigatus is the most prevalent saprophytic fungi
and can cause severe invasive aspergillosis in immunocompromised
individuals. For infection of A. fumigatus, the small
hydrophobic conidia have been shown to play a dominant
role. In this study, we found that deletion of erg5, a C-22 sterol
desaturase gene which function in the last two steps of ergosterol
biosynthesis, was sufficient to block ergosterol biosynthesis
and conidiation. The deletion phenotype was further
verified by a conditional expression strain of erg5 using the
inducible tet-on system. Strikingly, erg5 mutant displays increased
susceptibility to antifungal azoles itraconazole. RNA
sequencing analysis showed that erg5 deficiency resulted in
changes in transcription mainly related to lipid, carbohydrate,
and amino acid metabolism. Genes encoding ergosterol biosynthesis-
related enzymes were found to be up-regulated in
erg5 null mutants. However, genes involved in asexual development,
including upstream regulators, melanin biosynthesis
enzymes, heterotrimeric G proteins, and MAPK signaling,
were down-regulated to various degrees. Furthermore, metabolomic
study revealed that erg5 deficiency also resulted in
altered lipid and amino acid metabolism, which was consistent
with our transcriptomics analysis. Collectively, our study
established a link between ergosterol biosynthesis and asexual
development at the transcriptomics and metabolomics level
in A. fumigatus.
The phylogenetic study of Penicilium chrysogenum was performed based on amino acid sequence comparison of chitin synthase. Phylogenetic trees were constructed with the deduced amino acid sequences of the highly conserved region of chitin synthase gene fragments amplified by PCR. The BlasP similarity search and the bootstrap analysis of the deduced amino acid sequences of chitin synthase from P. chrysogenum with those form other fungi showed a close evolutionary relationship of Penicillium to ascomycetous fungi, especially to genus Aspergilus. The result from bootstrap analysis of the deduced amino acid sequences of the Class II chitin synthase from ascomyceteous fungi supported the usefulness of the Class II chitin synthease for phylogenetic study of filamentous fungi.
Class III chitin synthases in filamentous fungi are important for hyphal growth and differentiation of several filamentous fungi. A genomic clone containing the full gene encoding Chs4, a class III chitin synthase in Penicillium chrysogenum, was cloned by PCR screening and colony hybridization from the genomic library. Nucleotide sequence analysis and transcript mapping of chs4 revealed an open reading frame (ORF) that consisted of 5 exons and 4 introns and encoded a putative protein of 915 amino acids. Nucleotide sequence analysis of the 5?lanking region of the ORF revealed a potential TATA box and several binding sites for transcription activators. The putative transcription initiation site at ?6 position was identified by primer extension and the expression of the chs4 during the vegetative growth was confirmed by Northern blot analysis. Amino acid sequence analysis of the Chs4 revealed at least 5 transmembrane helices and several sites for post-translational modifications. Comparison of the amino acid sequence of Chs4 with those of other fungi showed a close relationship between P. chr ysogenum and genus Aspergillus.