Extension of O-linked mannosylation in the Golgi is critical for cell wall integrity signaling and interaction with host cells in Cryptococcus neoformans pathogenesis

Purpose: To identify the downstream gene sets regulated by Mpk1p in C. neoformans normal and Tunicamycin-treated conditions, we compared the RNA-seq based transcriptional profiles. Methods: Total RNA was isolated according to the RNeasy Mini Kit (Qiagen) and all procedure for RNA sequencing were conducted by ebiogen. The mRNA was isolated by using the Poly(A) RNA Selection Kit (LEXOGEN) and isolated mRNAs were used for the cDNA synthesis. Results: Transcriptome analysis of the wild-type strain revealed that tunicamycin significantly increased expression of a subset of calcineurin-Crz1 pathway related genes , but most of which were not upregulated in O-glycan mutant strain. Conclusion: Our results support the essential role of O-glycan in maintaining cell wall integrity upon stress conditions of C. neoformans. mRNA profiles of cells grown at normal and tunicamycin-treated conditions were generated by RNA-seq using NovaSeq 6000 (Illumina).