Differential expression of sialylglycoconjugates and sialidase activity in distinct morphological stages of Fonsecaea pedrosoi.
The expression of sialoglycoconjugates in Fonsecaea pedrosoi conidia, mycelia, and sclerotic cells was analyzed using influenza A and C virus strains, sialidase treatment, and lectin binding. Conidium and mycelium whole cells were recognized by Limax flavus (LFA), Maackia amurensis (MAA), and Sambucus nigra (SNA) lectins, denoting the presence of surface sialoglycoconjugates containing alpha 2,3- and alpha 2,6-sialylgalactosyl sequences. Sialidase-treated conidia reacted more intensively with peanut agglutinin (PNA), confirming the occurrence of sialyl-galactosyl linkages. Conidial cells agglutinated in the presence of influenza A and C virus strains, which confirmed the results obtained from lectin-binding experiments and revealed the presence of sialoglycoconjugates bearing 9-O-acetyl-N-acetylneuraminic acid (Neu5,9Ac(2)) surface structures. Western blotting analysis with peroxidase-labeled LFA demonstrated the occurrence of sialylglycoproteins in protein extracts from conidia and mycelia, with molecular masses corresponding to 56 and 40 kDa. An additional band of 77 kDa was detected in conidial extracts, suggesting an association between sialic acid expression and morphogenesis. Synthesis of sialic acids was correlated with sialidase expression, since both conidial and mycelial morphological stages presented secreted and cell-associated enzyme activity. Sialoglycoconjugates were not detected in F. pedrosoi sclerotic cells from in vitro and in vivo sources, which also do not express sialidase activity. The surface sialyl residues in F. pedrosoi are apparently involved in the fungal interaction with immune effector cells, since sialidase-treated conidia were less resistant to phagocytosis by human neutrophils from healthy individuals. These findings suggest that sialic acid expression in F. pedrosoi varies according to the morphological transition and may protect infecting propagules against immune destruction by host cells.
Alviano
Daniela S
DS
Instituto de Microbiologia Professor Paulo de Góes, Universidade Federal do Rio de Janeiro, Centro de Ciências da Saúde-Bloco I, Ilha do Fundão, 21941-590 Rio de Janeiro, RJ, Brazil.
Rodrigues
Marcio L
ML
Almeida
Catia A
CA
Santos
André L S
AL
Couceiro
José N S S
JN
Soares
Rosangela M A
RM
Travassos
Luiz R
LR
Alviano
Celuta S
CS
D001203:Ascomycota / Q000737:chemistry / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal; D006001:Glycoconjugates / Q000378:metabolism*; D029521:Alphainfluenzavirus / Q000378:metabolism; D009439:Neuraminidase / Q000737:chemistry / Q000378:metabolism*; D010587:Phagocytosis; D012794:Sialic Acids / Q000378:metabolism*
D006001:Glycoconjugates; D012794:Sialic Acids; D009439:Neuraminidase