Thomas, P.W., Wyckoff, E.E., Pishko, E.J., Yu, J.J., Kirkland, T.N., Cole, G.T. 1997. The hsp60 gene of the human pathogenic fungus Coccidioides immitis encodes a T-cell reactive protein. Gene. 199(1-2):83-91.
A heat shock protein-encoding gene (hsp60) from the human
respiratory fungal pathogen, Coccidioides immitis (Ci), was cloned, sequenced,
chromosome-mapped, expressed and immunolocalized in parasitic cells. Both the
genomic and cDNA sequences are presented. The transcription start point and poly
(A) addition site were confirmed. The hsp60 gene contains two introns and a
1782-bp ORF which translates a 594-amino acid (aa) protein of 62.4 kDa and pI of
5.6. The translated protein revealed two potential N-glycosylation sites. The
deduced HSP60 showed 78-83% aa sequence similarity to reported fungal HSP60
proteins. The hsp60 gene was mapped to chromosome III of Ci and was shown to be
a single copy gene by Southern and Northern hybridization. Expression of a
1737-bp cDNA fragment of the hsp60 gene in E. coli resulted in production of a
recombinant protein. Amino acid sequence analysis of the recombinant protein
confirmed that it was encoded by the Ci hsp60 gene. Antiserum raised in mice
against the isolated recombinant protein immunolocalized HSP60 in the cytoplasm
and wall of parasitic cells of Ci. The recombinant HSP60 was used to immunize
BALB/c mice and was shown to induce proliferation of T cells isolated from lymph
nodes of these animals. The hsp60 gene of Ci is the first reported heat-shock
protein gene of this human pathogen.