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. 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.
Department of Microbiology, University of Rochester School of Medicine and Dentistry, NY 14642, USA. Haid@uhura.cc.rochester.edu
C G
CG
Haidaris
O F
OF
Medzihradsky
F
F
Gigliotti
P J
PJ
Simpson-Haidaris
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017124:Conserved Sequence; D018076:DNA, Complementary; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics*; D015723:Gene Library; D005800:Genes, Fungal; D006801:Humans; D008168:Lung / Q000382:microbiology; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000235:genetics*; D051379:Mice; D016513:Mice, SCID; D008969:Molecular Sequence Data; D011010:Pneumocystis / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D011020:Pneumonia, Pneumocystis / Q000382:microbiology*; D051381:Rats; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
D018076:DNA, Complementary; D005656:Fungal Proteins; D008562:Membrane Glycoproteins; C077556:GPA protein, Pneumocystis carinii sp.