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Fusobacterium nucleatum Apoptosis-inducing Outer Membrane Protein

¹ Molecular Biology Institute,
² Department of Microbiology, Immunology and Molecular Genetics, and
³ Department of Oral Biology and Oral Medicine, Dental Research Institute, Section of Periodontics, UCLA School of Dentistry, 10833 Le Conte Ave., Los Angeles, CA 90095-1668;

View larger version (33K): [in this window] [in a new window]   Figure 1. Structural and transcriptional analyses of wild-type and aimless1 mutant strains. (A) Schematic illustration of mutagenesis: organization of aim1 and flanking genes on the F. nucleatum ATCC 23726 wild-type and aimless1 mutant chromosome. Arrows indicate PCR primers used for construction and analysis of the aim1 mutant; arrowheads indicate real-time PCR primers; black shaded area indicates intergenic region; bracket indicates location of autotransporter domain. Full-length aim1 (5625 bases) was submitted to GenBank and has been assigned accession number DQ020251. The Fig. is not to scale. (B) PCR analysis confirming insertion into aim1. PCR analysis of chromosomal DNA from ATCC 23726 and the aimless1 mutant. Plasmid pIP-aim1, the construct used to create the inactivation in aim1, was used as the control DNA template. (C) Transcriptional analysis of aim1 and the downstream gene Fn2057. The relative RNA abundance of F. nucleatum wild-type and aimless1 was determined by real-time PCR. Results are expressed as relative means (mutant/wt) ± SEMs (error bars) of triplicate experiments.

View larger version (12K): [in this window] [in a new window]   Figure 2. Induction of apoptosis in Jurkat cells after incubation with F. nucleatum wild-type or with aimless1. Jurkat cells were incubated with wild-type or aimless1 for 18 hrs before assay (n = 3). Uninfected Jurkat cells were incubated alone as a control. Values shown are relative to wild-type apoptosis levels after background subtraction. Error bars represent SD. **p < 0.0001.

Journal of Dental Research, Vol. 84, No. 8, 700-704 (2005)
DOI: 10.1177/154405910508400803


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