Molecular characterization of mouse Pneumocystis carinii surface glycoprotein A

Since the mouse offers an easily manipulated experimental animal model for the study of the immunopathogenesis of pneumonia caused by the opportunist Pneumocystis carinii, we cloned and characterized cDNAs encoding an abundant, immunogenic surface antigen termed glycoprotein A (gpA) from mouse P. ca...

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Bibliographic Details
Published in:DNA research 1998-04, Vol.5 (2), p.77-85
Main Authors: Haidaris, C G, Medzihradsky, O F, Gigliotti, F, Simpson-Haidaris, P J
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Base Sequence
Conserved Sequence
DNA, Complementary
Fungal Proteins - chemistry
Fungal Proteins - genetics
Gene Library
Genes, Fungal
Humans
Lung - microbiology
Membrane Glycoproteins - chemistry
Membrane Glycoproteins - genetics
Mice
Mice, SCID
Molecular Sequence Data
Pneumocystis - chemistry
Pneumocystis - genetics
Pneumocystis - isolation & purification
Pneumonia, Pneumocystis - microbiology
Rats
Sequence Alignment
Sequence Analysis, DNA
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Summary:Since the mouse offers an easily manipulated experimental animal model for the study of the immunopathogenesis of pneumonia caused by the opportunist Pneumocystis carinii, we cloned and characterized cDNAs encoding an abundant, immunogenic surface antigen termed glycoprotein A (gpA) from mouse P. carinii. A cDNA library was constructed in bacteriophage lambda gt11 from P. carinii-infected mouse lung poly(A+) RNA. Using a nucleic acid probe derived from a conserved region of the mouse P. carinii gpA structural gene, cDNAs encoding gpA were identified. A composite full-length gpA coding sequence was assembled from two overlapping cDNA clones. A DNA element homologous to the rat P. carinii upstream conserved sequence (UCS) was identified at the 5' end of several of the mouse P. carinii gpA cDNA clones, just upstream of the sequences encoding gpA structural gene isoforms. Using primer extension analysis, two neighboring putative transcriptional start sites were located on UCS-gpA mRNAs approximately 25 and 30 nt, respectively, upstream of the most 5' gpA cDNA clone isolated, suggesting a 5' UCS of 489 or 494 nucleotides in mouse P. carinii gpA. A comparative alignment of the composite mouse P. carinii gpA deduced amino acid sequence with gpA homologs from rat, human and ferret P. carinii demonstrated 156 identical residues, including 46 cysteines, further supporting the hypothesis for conserved secondary structure, as well as function, for gpA from all P. carinii.
ISSN:1340-2838
DOI:10.1093/dnares/5.2.77