Cloning and expression of the gene which encodes a tube precipitin antigen and wall-associated beta-glucosidase of Coccidioides immitis.
We report the structure and expression of the Coccidioides immitis BGL2 gene which encodes a previously characterized 120-kDa glycoprotein of this fungal respiratory pathogen. The glycoprotein is recognized by immunoglobulin M tube precipitin (TP) antibody present in sera of patients with coccidioidomycosis, a reaction which has been used for serodiagnosis of early coccidioidal infection. The deduced amino acid sequence of BGL2 shows 12 potential N glycosylation sites and numerous serine-threonine-rich regions which could function as sites for O glycosylation. In addition, the protein sequence includes a domain which is characteristic of family 3 glycosyl hydrolases. Earlier biochemical studies of the purified 120-kDa TP antigen revealed that it functions as a beta-glucosidase (EC 3.2.1.21). Its amino acid sequence shows high homology to several other reported fungal beta-glucosidases which are members of the family 3 glycosyl hydrolases. Results of previous studies have also suggested that the 120-kDa beta-glucosidase participates in wall modification during differentiation of the parasitic cells (spherules) of C. immitis. In this study we showed that expression of the BGL2 gene is elevated during isotropic growth of spherules and the peak of wall-associated BGL2 enzyme activity correlates with this same phase of parasitic cell differentiation. These data support our hypothesis that the 120-kDa beta-glucosidase plays a morphogenetic role in the parasitic cycle of C. immitis.
Hung
C Y
CY
Department of Microbiology and Immunology, Medical College of Ohio, Toledo, Ohio 43614-5806, USA.
Yu
J J
JJ
Lehmann
P F
PF
Cole
G T
GT
D000595:Amino Acid Sequence; D000908:Antibodies, Fungal / Q000276:immunology*; D000946:Antigens, Fungal / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003045:Coccidioides / Q000235:genetics* / Q000276:immunology; D005800:Genes, Fungal; D007075:Immunoglobulin M / Q000276:immunology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011233:Precipitin Tests; D020133:Reverse Transcriptase Polymerase Chain Reaction; D001617:beta-Glucosidase / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification
D000908:Antibodies, Fungal; D000946:Antigens, Fungal; D007075:Immunoglobulin M; D001617:beta-Glucosidase