DNA sequencing of the gene encoding a bacterial superantigen, Yersinia pseudotuberculosis-derived mitogen (YPM), and characterization of the gene product, cloned YPM.
Previously, we found a novel bacterial superantigen from Yersinia pseudotuberculosis, designated Y. pseudotuberculosis-derived mitogen (YPM). In the present study, we analyzed the DNA sequence of the gene encoding YPM. The YPM gene was cloned into a plasmid vector pMW119 and expressed in Escherichia coli DH10B. Like the native YPM, the cloned YPM required the expression of MHC class II molecules on accessory cells in the induction of IL-2 production by human T cells. TCR-V beta repertoire of human T cells reactive with the cloned YPM was V beta 3, V beta 9, V beta 13.1, and V beta 13.2. This repertoire is the same as that of T cells reactive with the native YPM. These results indicate that the cloned YPM expressed in E. coli is identical to the native YPM. Sequencing of the YPM gene revealed that the gene contained an open reading frame of 456 base pairs encoding a precursor form of 151 amino acid residues with m.w. 16,679 that is processed into a mature form of 131 amino acid residues with m.w. 14,529. Homology analysis revealed that the homology of amino acid sequence is quite low among YPM and other well known bacterial superantigens. We designated the gene encoding YPM as ypm.
Miyoshi-Akiyama
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Department of Microbiology and Immunology, Tokyo Women's Medical College, Japan.
Abe
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A
Kato
H
H
Kawahara
K
K
Narimatsu
H
H
Uchiyama
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D015153:Blotting, Western; D002478:Cells, Cultured; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D015698:Genomic Library; D006801:Humans; D007739:L Cells; D051379:Mice; D008934:Mitogens / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics; D011994:Recombinant Proteins; D018089:Superantigens / Q000235:genetics* / Q000276:immunology; D015011:Yersinia pseudotuberculosis / Q000235:genetics* / Q000276:immunology
D001426:Bacterial Proteins; D008934:Mitogens; D016693:Receptors, Antigen, T-Cell, alpha-beta; D011994:Recombinant Proteins; D018089:Superantigens; C093313:Yersinia pseudotuberculosis-derived mitogen