Legionella pneumophila htpAB heat shock operon: nucleotide sequence and expression of the 60-kilodalton antigen in L. pneumophila-infected HeLa cells.
A 60-kilodalton (kDa) immunodominant antigen of Legionella pneumophila is a heat shock protein (HSP) of the GroEL class of HSPs. The gene (htpB) coding the 60-kDa protein was localized to a 3.2-kilobase DNA fragment of L. pneumophila cloned into pUC19 (pSH16) (P. S. Hoffman, C. A. Butler, and F. D. Quinn, Infect. Immun. 57:1731-1739, 1989). The nucleotide sequence of the DNA fragment cloned into M13 confirmed two open reading frames, htpA and htpB, that code for proteins of 96 and 548 amino acids, respectively. A consensus heat shock promoter sequence upstream of the start of htpA was identified, and no obvious promoter sequences were detected upstream of htpB. Amino acid sequence comparison studies revealed that the L. pneumophila HtpB protein exhibited 76% homology with the 65-kDa protein of Mycobacterium tuberculosis and 85% homology with both GroEL of Escherichia coli and HtpB of Coxiella burnetii. A comparison of the amino acid sequences among these proteins revealed several regions of nearly absolute sequence conservation, with the variable regions occurring in common areas. The purified L. pneumophila 60-kDa protein was antigenic for human T lymphocytes. Indirect fluorescent antibody studies indicated that the 60-kDa protein may be located in the periplasm or expressed on the surface by intracellular bacteria, suggesting that a stress-related mechanism may be involved in the expression of this immunodominant antigen.
Hoffman
P S
PS
Department of Microbiology and Immunology, University of Tennessee, Memphis 38163.
Houston
L
L
Butler
C A
CA
D000595:Amino Acid Sequence; D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D000954:Antigens, Surface / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D001483:Base Sequence; D004269:DNA, Bacterial; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D006801:Humans; D007875:Legionella / Q000235:genetics*; D008213:Lymphocyte Activation / Q000276:immunology; D008969:Molecular Sequence Data; D009876:Operon; D012689:Sequence Homology, Nucleic Acid
D000942:Antigens, Bacterial; D000954:Antigens, Surface; D004269:DNA, Bacterial; D006360:Heat-Shock Proteins