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Raw sequence reads of Candida albicans Hap31 and Hap32, two subunits of the CCAAT binding complex

ABSTRACT: The opportunistic fungal pathogen Candida albicans has recently impacted a substantial portion of the population. The highly conserved CCAAT-binding complex (CBC), also known as the Hap complex in yeast, plays a critical role in multiple cellular functions. Additionally, the cell wall of this pathogen serves as an effective barrier against environmental stresses and undergoes continuous renewal through a complex regulatory network. This network is tightly controlled by numerous cell signaling pathways and transcription factors. Mutants with individual deletions of components of the Hap complex were generated and compared to the wild-type strain to investigate the regulatory role of the CBC in cell wall maintenance and biofilm formation. Additionally, RNA sequencing was performed to uncover novel insights into the regulatory pathways associated with the CBC. Deletion of CBC components significantly impacts the cell wall structure and composition, leading to an imbalance in the cell wall carbohydrate profile, as well as cell wall unmasking, which triggers the activation of the Cell Wall Integrity (CWI) signaling pathway. These alterations subsequently affect the ability to form biofilms on polystyrene surfaces. Notably, the small GTPase Rhb1 is implicated in CBC-mediated CWI maintenance. Moreover, RNA sequencing of the expression profiles of the two Hap3 paralogs suggests their involvement in rRNA processing, ribosomal biogenesis, carbohydrate transport and metabolism, and amino acid biosynthesis, all contributing to cell wall maintenance. Our findings provide novel insights into the molecular pathways that govern cell wall biogenesis. The discovery of this regulatory mechanism is crucial for understanding the relationship between the highly conserved CBC and cell wall maintenance in C. albicans.

ORGANISM(S): Candida albicans SC5314

PROVIDER: GSE293407 | GEO | 2025/09/15

REPOSITORIES: GEO

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Project description:As a dynamic structure with a barrier between the cell and the outside world, maintenance of cell wall integrity (CWI) is important for fungal growth, development and pathogenicity processes. Here we characterized a MADS-box transcription factor RlmA (AoRlmA) downstream of the CWI regulatory pathway in the nematode-trapping fungus Arthrobotrys oligospora. Deletion of AorlmA caused a reduction in mycelial growth and the number of nuclei, and significant downregulation of transcript levels of genes related to nucleus synthesis and DNA damage repair, such as rad3, spo11, and rad25. Meanwhile, the ΔAorlmA mutant strains showed a significant reduction in spore production compared with the wild-type (WT) strain, and the transcript levels of sporulation-related genes was downregulated in the ΔAorlmA mutant during the early and middle stages of condiation, such as flbA, medA, and vosA. Meanwhile, the mycelial cell wall of the ΔAorlmA mutant strain showed breakage. Also, the transcript levels of genes related to cell wall synthesis were significantly down-regulated in the mutant strain compared to the WT strain, and the mutant strain was more sensitive to stresses of cell wall synthesis disruptors, oxidative stress and osmotic stress than the WT strain. Compared with the WT strain, the ΔAorlmA mutant strain not only had a reduced number of traps and nematicidal ability, but also, the shape of the traps of the mutants has also changed. In addition, In addition, AoRlmA also regulates autophagy and endocytosis. Transcriptome analysis of ΔAorlmA mutant strains showed that AorlmA regulates redox, cell wall synthesis, DNA replication and damage repair, and pathogenic processes. Metabolomic analysis showed that AorlmA is involved in the biosynthesis of secondary metabolites of A. oligospora. To summarise, our data suggest that AorlmA is involved in growth, sporulation, spore germination, maintenance of cell wall integrity, DNA replication and damage repair, pathogenesis, autophagy and secondary metabolite biosynthesis processes.

Project description:The Neurospora crassa GUL-1 is part of the COT-1 pathway, which plays key roles in regulating polar hyphal growth and cell wall remodeling. We show that GUL-1 is a bona fide mRNA binding protein (RBP) that can associate with 828 “core” mRNA species. When cell wall integrity (CWI) is challenged, the repertoire of associated RNA species increases up to 2628 mRNAs (including its own transcript). GUL-1 can bind mRNAs of genes related to translation, cell wall remodeling, circadian clock, endoplasmatic reticulum (ER), as well as CWI and osmoregulatory response MAPK pathway components. GUL-1 interacts with over 100 different proteins, including stress granule proteins, components of the translational and cell wall remodeling machinery, as well as ER components and those of the MAPK, COT-1 and STRIPAK complexes. Several additional RBPs were also shown to physically interact with GUL-1. We demonstrate that under stress conditions GUL-1 can localize to the ER and that GUL-1 affects the CWI pathway as is evident via altered phosphorylation levels of MAK-1, interaction with mak-1 transcript and involvement in the expression level of the transcription factor adv-1. Taken together, we suggest that the RBP GUL-1 functions in multiple cellular processes, including the regulation of cell wall remodeling. The latter is mechanistically concerted with the MAK-1 pathway in a stress-related manner. Methods: In this study we show that GUL-1 is a bona fide mRNA binding protein (RBP) that associates with different mRNAs, including its own transcript, as determined by RNA antisense purification and RNA immunoprecipitation experiments. Results: We demonstrate that GUL-1 may physically interact with multiple proteins included translation processes, cell wall remodeling and cell wall integrity (CWI). We show that GUL-1 affects the CWI pathway as is evident via altered phosphorylation levels of MAK-1, interaction with mak-1 transcript and involvement in the expression level of the transcription factor adv-1. Conclusions:We suggest that the RBP GUL-1 functions in the regulation of cell wall remodeling in concert with the MAK-1 pathway in a stress-related manner.

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Raw sequence reads of Candida albicans Hap31 and Hap32, two subunits of the CCAAT binding complex

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