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Lack of chitin synthase genes impacts capsular architecture and cellular physiology in Cryptococcus neoformans

作者：    Jessica Rodrigues^a, Caroline L. Ramos^a,  Susana Frases^b, Rodrigo M. da C. Godinho^a, Fernanda L. Fonseca^c, Marcio L. Rodrigues^{a, d}

    a Instituto de Microbiologia Paulo de Góes, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil

    b Instituto de Biofísica Carlos Chagas Filho, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil

    c Centro de Desenvolvimento Tecnológico em Saúde (CDTS) da Fundação Oswaldo Cruz (Fiocruz), Rio de Janeiro, Brazil

    d Instituto Carlos Chagas (ICC), Fundação Oswaldo Cruz (Fiocruz), Curitiba, PR, Brazil

摘要：Cryptococcus neoformans mutants lacking each of the eight putative chitin synthase genes (CHS) have been previously generated. However, it is still unclear how deletion of chitin synthase genes affects the cryptococcal capsule. Since the connections between chitin metabolism and capsular polysaccharides in C. neoformans are numerous, we analyzed the effects of deletion of CHS genes on capsular and capsule-related structures of C. neoformans. CHS deletion affected capsular morphology in multiple ways, as determined by scanning electron microscopy and immunofluorescence analysis. Molecular diameter, serological reactivity and export of capsular polysaccharide were also affected in most of the chsΔ mutants, but the most prominent alterations were observed in the chs3Δ strain. C. neoformans cells lacking CHS genes also had altered formation of extracellular vesicles and variable chitinase activity under stress conditions. These results reveal previously unknown functions of CHS genes that greatly impact the physiology of C. neoformans.