Trial Information

Novel Protein Regulator of Tumor Suppressor ARF & NF-κB

OBJECTIVES:

- Define the function and mechanism of LZAP in regulating ARF.

- Determine the mechanism and biological consequences of LZAP inhibition of NF-κB.

- Determine if LZAP has tumor suppressor activity by conditional targeting of LZAP in
mice.

OUTLINE: Using molecular laboratory techniques, this study examines the biochemical
mechanisms by which LZAP activates transcriptional, tumor suppressive activity (both
p53-dependent and p53-independent) of ARF and inhibits transcriptional, tumorigenic activity
of NF-kB. LZAP regulation of newly identified ARF activities, such as S-phase delay and
ARF-mediated B23 degradation are also evaluated. LZAP's tumor suppressive activity is
assessed in vivo using a conditional knockout vector to target LZAP in mice and to observe
for spontaneous and induced tumor formation. Laboratory analyses used to determine study
endpoints include standard recombinant DNA, recombinant protein expression and purification,
cell culture and transfection, cell labeling, reporter assays, flow cytometry, yeast-two
hybrid, immunoprecipitation, immunoblotting, immunofluorescence, TUNEL assay, ELISA assay,
Southern blotting, and protein and gene expression.